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89,553
Quantification of p62-positive inclusions
4
dx.doi.org/10.17504/protocols.io.8epv5x33ng1b/v1
https://www.protocols.io/view/quantification-of-p62-positive-inclusions-c3prymm6
Núria Peñuelas
TITLE: Quantification of p62-positive inclusions AUTHORS: Núria Peñuelas [DESCRIPTION] Quantification of p62-positive inclusions in the rodent brain [STEPS] 1. Scan the 5um sections immunostained with guineapig anti-p62 (1:500, Progen #GP62-C), mouse anti-α-synuclein (1:500, BD Biosciences #610786) and TH (1:1000, Ca...
["Scan the 5um sections immunostained with guineapig anti-p62 (1:500, Progen #GP62-C), mouse anti-α-synuclein (1:500, BD Biosciences #610786) and TH (1:1000, Calbiochem #657012) using ZEISS LSM 980 with Airyscan 2.", "Visualize the images with ZEN 2011 software (Zeiss, Germany) and determine one representative coronal ...
78,975
Efficacy and safety of probiotics in the treatment of irritable bowel syndrome: a systematic review and meta-analysis of randomised clinical trials using ROME IV criteria
1
null
https://www.protocols.io/view/efficacy-and-safety-of-probiotics-in-the-treatment-crc7v2zn
Giorgos Konstantis, Efstathiou Stylianos, Chryssa Pourzitaki, Kitsikidou Elisavet, Germanidis Georgios, Chourdakis Michail
TITLE: Efficacy and safety of probiotics in the treatment of irritable bowel syndrome: a systematic review and meta-analysis of randomised clinical trials using ROME IV criteria AUTHORS: Giorgos Konstantis, Efstathiou Stylianos, Chryssa Pourzitaki, Kitsikidou Elisavet, Germanidis Georgios, Chourdakis Michail [DESCRIPT...
["Systematic review\n\nReview title.\n\n\nEfficacy and safety of probiotics in the treatment of irritable bowel syndrome: a systematic review and meta-analysis of randomised clinical trials using ROME IV criteria\n\nAnticipated or actual start date.\n\n\n\n01 October 2021 \n\n\nAnticipated completion date.\n\n\nNovembe...
31,407
AAV Production in HEK293T Cells
null
dx.doi.org/10.17504/protocols.io.bawpifdn
null
Addgene the nonprofit plasmid repository
TITLE: AAV Production in HEK293T Cells AUTHORS: Addgene the nonprofit plasmid repository [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol is used for AAV production in HEK293T cells. To see the full abstract and additional resources, visit the </span><a href="https://www.addgene.o...
["[Procedure]\nTrypsinize and resuspend the HEK293T packaging cells from 2 x T-150 flasks. Cells should be at ~80% confluence. For each T-150 flask:Aspirate culture media and rinse once with of PBS.Aspirate PBS and add of 0.05% Trypsin/EDTA. Wait ~.Neutralize trypsin by adding of HI-FBS.\n10 ml\n4 ml\n4 ml", "[Proce...
53,590
Quadro de trabalho para Revisões Sistemáticas e Quantitativas da Literatura (RSQL)
1
dx.doi.org/10.17504/protocols.io.byjwpupe
https://www.protocols.io/view/quadro-de-trabalho-para-revis-es-sistem-ticas-e-qu-byjwpupe
Daniel Vartanian
TITLE: Quadro de trabalho para Revisões Sistemáticas e Quantitativas da Literatura (RSQL) AUTHORS: Daniel Vartanian [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Este protocolo foi criado para o programa de Revisões Sistemáticas e Quantitativas da Literatura (RSQL) científica do Grupo Interd...
["[Método Pickering & Byrne]\nConheça o método de Revisões Sistemáticas e Quantitativas da Literatura (RSQL) de Pickering e Byrne\nEste programa utiliza o método desenvolvido por Catherine Pickering e Jason Byrne. A primeira coisa que você precisa fazer é obter um bom conhecimento sobre ele.", "[Método Pickering & Byrn...
48,184
Antioxidant activity by reduced glutathione (GSH) assay: in vitro protocol
6
dx.doi.org/10.17504/protocols.io.btaynifw
https://www.protocols.io/view/antioxidant-activity-by-reduced-glutathione-gsh-as-btaynifw
Adrieli Sachett, Matheus Gallas-Lopes, Greicy M M Conterato, Ana Herrmann, Angelo Piato
TITLE: Antioxidant activity by reduced glutathione (GSH) assay: in vitro protocol AUTHORS: Adrieli Sachett, Matheus Gallas-Lopes, Greicy M M Conterato, Ana Herrmann, Angelo Piato [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left"><span>Considering the ro...
["[Preparing the reagents]\nThe first step is to prepare the reagents to be used in this protocol;", "[Preparing the reagents]\nPotassium phosphate buffer : 1.1.1 Weigh of monobasic potassium phosphate (KH2PO4) in a beaker of appropriate size; 1.1.2 Dissolve the salt with of ultrapure water; 1.1.3 Transfer t...
10,022
Validity of daily self-pulse palpation over two weeks for screening for atrial fibrillation among patients 65 years of age and older seeking primary care
null
dx.doi.org/10.17504/protocols.io.m2ec8be
null
Faris Ghazal
TITLE: Validity of daily self-pulse palpation over two weeks for screening for atrial fibrillation among patients 65 years of age and older seeking primary care AUTHORS: Faris Ghazal [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left"><span style = "font-...
["Design and Selection", "Design: Cross sectional study.", "Study population: It was planned to include patients who, for any reason, were seeking care in the primary health care and who were 65 years of age or older. Patients with previously known atrial fibrillation or ongoing anticoagualtion treatment were exluded."...
51,287
Cross-cultural adaptation and validity of the Italian version of the Anterior Knee Pain Scale: a prospective multi-centre study
1
dx.doi.org/10.17504/protocols.io.bwbxpapn
https://www.protocols.io/view/cross-cultural-adaptation-and-validity-of-the-ital-bwbxpapn
Jacopo E. Rocchi, Pasquale Alessio Sauchelli, Sebastiano Nutarelli, Lorenzo Rum, Riccardo Ciatti
TITLE: Cross-cultural adaptation and validity of the Italian version of the Anterior Knee Pain Scale: a prospective multi-centre study AUTHORS: Jacopo E. Rocchi, Pasquale Alessio Sauchelli, Sebastiano Nutarelli, Lorenzo Rum, Riccardo Ciatti [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Purpose: va...
[]
48,346
Single-cell suspension preparation from Human bronchial biopsies to perform scRNA-sequencing using 10x chromium
4
null
https://www.protocols.io/view/single-cell-suspension-preparation-from-human-bron-btf2njqe
Martijn Nawijn, Leonie Apperloo
TITLE: Single-cell suspension preparation from Human bronchial biopsies to perform scRNA-sequencing using 10x chromium AUTHORS: Martijn Nawijn, Leonie Apperloo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol enables the dissociation of human bronchial biopsies into a single-cell suspen...
["[Before you start]\nAll steps should be performed in a sterile environment. Prepare all the buffers and reagents at room temperature as described in the \"Materials\" section.", "[Before you start]\nFreshly prepare the tissue dissociation solution.Dilute the 10x RBCs lysis buffer to 1x.", "[Preparation of single-cel...
67,451
Wo Sie Diaetovita (Höhle Der Löwen und Dietavita Erfahrungen) jetzt auf der offiziellen Website kaufen können!
1
dx.doi.org/10.17504/protocols.io.14egn7d8mv5d/v1
https://www.protocols.io/view/wo-sie-diaetovita-h-hle-der-l-wen-und-dietavita-er-cd43s8yn
forexcarter
TITLE: Wo Sie Diaetovita (Höhle Der Löwen und Dietavita Erfahrungen) jetzt auf der offiziellen Website kaufen können! AUTHORS: forexcarter [DESCRIPTION] Produktname: Diaetovita (Höhle Der LöwenundDietavita Erfahrungen) Nebenwirkung: Keine größeren Nebenwirkungen bekannt Erwartete Ergebnisse: In 2 – 3 Monaten Bewe...
["[Wo Sie Diaetovita (Höhle Der Löwen und Dietavita Erfahrungen) jetzt auf der offiziellen Website kaufen können!]"]
39,339
Characterization
1
null
https://www.protocols.io/view/characterization-binjkdcn
Hung Liang Pai
TITLE: Characterization AUTHORS: Hung Liang Pai [STEPS] ?. [In vitro protein synthesis] Add the reagents(on ice) for synthesis of mRFP into one well of the 384-well plate. Add the reagents from top to down. ABCD1OrderLocationReagentAmount21Upper-leftDNase/RNase free waterTill 30 μL32Upper-leftSolution A12μL43Lower-r...
["[In vitro protein synthesis]\nAdd the reagents(on ice) for synthesis of mRFP into one well of the 384-well plate. Add the reagents from top to down. ABCD1OrderLocationReagentAmount21Upper-leftDNase/RNase free waterTill 30 μL32Upper-leftSolution A12μL43Lower-rightSolution B9μL54Lower-rightRNase inhibitor1.2μL65Lower...
55,650
SWAP mouse genotyping
4
null
https://www.protocols.io/view/swap-mouse-genotyping-b2kaqcse
Peter W W Chomczynski, Kianna M Vires, Michal Rymascewski, Judith A. Heiny
TITLE: SWAP mouse genotyping AUTHORS: Peter W W Chomczynski, Kianna M Vires, Michal Rymascewski, Judith A. Heiny [DESCRIPTION] The highly-conserved, cardiotonic steroid (CTS) binding site (also termed ouabain binding site) on the primary α subunit of Na,K-ATPase (NKA) plays a receptor signaling role in a range o...
["[Tail clip digestion] Briefly vortex and centrifuge the tubes to pellet insoluble material. DNA will remain in the supernatant.", "[Tail clip digestion] Create a 1:100 dilution of each sample in a new, labelled 1.5 mL microcentrifuge tube. Add 198 µL of TE buffer (or ddH2O) to each tube, then add 2.0 µL of supernatan...
76,288
Immunocytochemistry
4
dx.doi.org/10.17504/protocols.io.5qpvor7pdv4o/v1
https://www.protocols.io/view/immunocytochemistry-cnq8vdzw
anita.adami
TITLE: Immunocytochemistry AUTHORS: anita.adami [DESCRIPTION] This protocol describes how to perform immunocytochemistry on 2D fixed cells [STEPS] SECTION: Fixing 1. The cells were washed three times with DPBS (GIBCO) and fixed for 10 minutes with 4% paraformaldehyde (Merck Millipore), followed by three more rinses ...
["[Fixing] The cells were washed three times with DPBS (GIBCO) and fixed for 10 minutes with 4% paraformaldehyde (Merck Millipore), followed by three more rinses with DPBS.", "[Blocking and permeabilisation] The cells were blocked for 60 min in blocking solution (KPBS 0.25% Triton X-100 (Fisher Scientific) and 5% norma...
71,022
Inductively coupled plasma mass spectrometry (ICP-MS)
6
dx.doi.org/10.17504/protocols.io.x54v9dj2pg3e/v1
https://www.protocols.io/view/inductively-coupled-plasma-mass-spectrometry-icp-m-chknt4ve
An.Huang
TITLE: Inductively coupled plasma mass spectrometry (ICP-MS) AUTHORS: An.Huang [DESCRIPTION] Inductively coupled plasma mass spectrometry (ICP-MS) can be used to detect metal elements existing in samples. Because we are not capable of conducting ICP-MS on our own, we must seek help from our advisors. This protocol is ...
["Filter all samples to be tested using 0.22μm syringe filters.", "(Optional) Dilute the samples to 0-20ppb using nitric acid (2%) if it contains too much desired elements more than the measuring range of equipment.", "Asking an assistant for help in conducting ICP-MS. Obtain a standardized curve before starting to mea...
null
null
null
dx.doi.org/10.17504/protocols.io.t5xeq7n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Oral cancer is one of the most significant cancers in the world; the most common of which is oral squamous cell carcinoma (OSCC) that makes up to 94% of oral malignances (Bernestein et al, 2013). Based on reports of the world Health Organization (WHO), OSCC occupied the sixth mo...
[]
95,972
BAF_Protocol_007 Solution Digest with Protein Precipitation
0
dx.doi.org/10.17504/protocols.io.bp2l6xjp5lqe/v1
https://www.protocols.io/view/baf-protocol-007-solution-digest-with-protein-prec-c9ycz7sw
Nicholas Sherman
TITLE: BAF_Protocol_007 Solution Digest with Protein Precipitation AUTHORS: Nicholas Sherman [DESCRIPTION] The protocol is a method to digest a protein mixture obtained from a cell lysis or tissue that is in solution. The protocol includes a precipitation step to clean up the proteins before digestion. The protein mix...
["[Solution Tissue Digest with Precipitation] Add 5X (volume to mg tissue) of lysis buffer (1% SDS, 100mM AmBiC) to tissue and transfer to Bead Mill 24 reinforced tube with 5 stainless steel balls.", "[Solution Tissue Digest with Precipitation] Lyse the cells with Fisher Bead Mill 24 (speed: 5m/s, time: 20 sec, number...
42,691
Dispensing agar into multiwell plates
1
dx.doi.org/10.17504/protocols.io.bmxbk7in
https://www.protocols.io/view/dispensing-agar-into-multiwell-plates-bmxbk7in
Ida Barlow
TITLE: Dispensing agar into multiwell plates AUTHORS: Ida Barlow [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Protocol for pouring agar into 96 well plates using Integra VIAFILL dispenser. Agar should be prepared in advance, and kept in 60</span><span style = "vertical-align:super;">o</span...
["[Configure Integra VIAFILL]\nPrepare a 250ml bottle of hot milliQ water in the microwave and keep in the waterbath along with the agar. The water is important to have on hand in case of tubing blockages.", "[Configure Integra VIAFILL]\nInsert large cassette into the machine", "[Configure Integra VIAFILL]\nConfigure X...
29,336
TotalSeq™-A Antibodies Protocol with 10x Single Cell 3&#39; Reagent Kit v2
null
dx.doi.org/10.17504/protocols.io.8vyhw7w
null
Sam Li
TITLE: TotalSeq™-A Antibodies Protocol with 10x Single Cell 3&#39; Reagent Kit v2 AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLege...
["[I) Cell staining for Drop-seq or 10x Genomics platforms]\nCarefully count all cells to ensure accurate quantitation.Make note of cell viability (>95%) and also include dead cells in the total cell count.If high cell death is observed, live cell enrichment (e.g. by Flow Cytometry) is recommended.", "[I) Cell staining...
95,641
Fecal sample collection
0
dx.doi.org/10.17504/protocols.io.3byl4qp22vo5/v1
https://www.protocols.io/view/fecal-sample-collection-c9mzz476
daniel.dautan daniel, Per Svenningsson
TITLE: Fecal sample collection AUTHORS: daniel.dautan daniel, Per Svenningsson [DESCRIPTION] Fecal sample collection from mice to use for gut microbiome sequencing analysis. [STEPS] 1. Set up the experiment by arranging clean cages without food, water or bedding. It is advisable to label each cage to avoid confusion ...
["Set up the experiment by arranging clean cages without food, water or bedding. It is advisable to label each cage to avoid confusion later on.", "In front of each cage, place a glass filled with dry ice and a 1.5ml Eppendorf tube to\nspeed up collection. Use plastic forceps each cage. Avoid using the same forceps to ...
75,598
Isolation of bacteria and fungi from cheese rind microbiomes
1
dx.doi.org/10.17504/protocols.io.rm7vzb7b4vx1/v1
https://www.protocols.click/view/isolation-of-bacteria-and-fungi-from-cheese-rind-m-cm3nu8me
Emily C.P. Weiss
TITLE: Isolation of bacteria and fungi from cheese rind microbiomes AUTHORS: Emily C.P. Weiss [DESCRIPTION] This protocol describes how to harvest and stock bacterial and fungal isolates from cheese rind microbial community samples. [STEPS] SECTION: Harvesting the cheese rind biofilm 2. Holding the cheese in one (glo...
["[Harvesting the cheese rind biofilm] Holding the cheese in one (gloved) hand and a sterile razor blade in the other, gently scrape the surface of the cheese to remove the rind biofilm. Avoid applying too much pressure, as you will start to dig into the paste. Scrape the rind biofilm into a weigh boat or other contain...
97,664
10x Protocols: Chromium Next GEM Single Cell 5&#39; -- University of Minnesota TMCsTMCs (CG000331 Rev E)
0
dx.doi.org/10.17504/protocols.io.5qpvokn89l4o/v1
https://www.protocols.io/view/10x-protocols-chromium-next-gem-single-cell-5-39-u-dbk82kzw
Laura J Niedernhofer, David A Bernlohr
TITLE: 10x Protocols: Chromium Next GEM Single Cell 5&#39; -- University of Minnesota TMCsTMCs (CG000331 Rev E) AUTHORS: Laura J Niedernhofer, David A Bernlohr [DESCRIPTION]  Following single-cell dissociation or nuclei isolation, the Next GEM 5' assay uses microfluidics to partition and assign cell- or nuclei-specifi...
["[Library Preparation]", "[FASTQ Generation] BCL data from Illumina sequencer is demultiplexed and converted into FASTQ format using bcl2fastq version 2.20.0", "[Tissue Preparation] Complete single cell or nuclei isolation prior to starting this protocol"]
74,646
0.1% BSA media prep and flow
4
dx.doi.org/10.17504/protocols.io.x54v9d7zpg3e/v1
https://www.protocols.io/view/0-1-bsa-media-prep-and-flow-ck5wuy7e
Olya Oppenheim
TITLE: 0.1% BSA media prep and flow AUTHORS: Olya Oppenheim [DESCRIPTION] Fetal Bovine Serum contains traces of ligands of the BMP pathway which affect the flow experiments with the ibidi perfusion system. replacing EGM2 with 2% with EGM2 with 0.1% BSA 4h prior to onset of flow (with same media) allows cells to respon...
["[BSA 0.1% media prep] 50 mL EGM2 without FBS\n667 µL BSA 7.5% solution\nfiltrate through 0.22um yellow filter.\nAdd to slides 4h before starting flow.", "[Optional] adding BMP9/10 to flow experiment\nin order to obtain a final 0.5 ng/ml , add 1.4ul BMP9/10 of 10 ug/ml initial concentration per perfusion unit (volume ...
34,133
unexpected perturbations while walking in virtual reality environment
1
dx.doi.org/10.17504/protocols.io.bdjvi4n6
https://www.protocols.io/view/unexpected-perturbations-while-walking-in-virtual-bdjvi4n6
Uri Rosenblum, Lotem Kribus-Shmiel, Gabi Zeilig, Yotam Bahat, Shani Kimel-Naor, Itshak Melzer, Meir Plotnik
TITLE: unexpected perturbations while walking in virtual reality environment AUTHORS: Uri Rosenblum, Lotem Kribus-Shmiel, Gabi Zeilig, Yotam Bahat, Shani Kimel-Naor, Itshak Melzer, Meir Plotnik [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Walking perturbations are a well-established tool to study...
["[Recruit Participants]\nIclusion CriteriaParticipants can be included in the study if they are:* Usually healthy* age 18-90Exclusion criteria Participants cannot be included in the study if they present the following conditions:* obesity (body mass index (kg/m2)> 30 [1]) * orthopedic condition affecting gait and bala...
88,153
Optimized QIAGEN DNeasy Blood &amp; Tissue kit Protocol for Environmental DNA Extraction
4
dx.doi.org/10.17504/protocols.io.8epv5xn66g1b/v1
https://www.protocols.io/view/optimized-qiagen-dneasy-blood-amp-tissue-kit-proto-c2bzyap6
Marion Chevrinais, Jade Larivière, Laury-Ann Dumoulin, Ariane Therien, Grégoire Cortial, Cloé Lepage, Eric Parent, Geneviève J. Parent
TITLE: Optimized QIAGEN DNeasy Blood &amp; Tissue kit Protocol for Environmental DNA Extraction AUTHORS: Marion Chevrinais, Jade Larivière, Laury-Ann Dumoulin, Ariane Therien, Grégoire Cortial, Cloé Lepage, Eric Parent, Geneviève J. Parent [DESCRIPTION] This document aims at providing a transparent method and detailin...
["[Filter Preparation] Install all the material on the benchtop including 2 mL microtubes with Lyse&Spin baskets for extraction, pre-identified 2 mL Eppendorf Safe-Lock for back up, tweezers, scissors, nacelles, waste beaker, microtube opener, and gloves.", "[Filter Preparation] Above a clean nacelle, take a filter wit...
41,694
COVID19 RTLAMP Assay
4
dx.doi.org/10.17504/protocols.io.bkx6kxre
https://www.protocols.io/view/covid19-rtlamp-assay-bkx6kxre
matt , Eugene Joseph, Arun Manoharan Arunprimediscoveriescom
TITLE: COVID19 RTLAMP Assay AUTHORS: matt , Eugene Joseph, Arun Manoharan Arunprimediscoveriescom [STEPS] ?. [Sample Purification / Concentration] Preheat the Heat Block or Water Bath ?. [RT-LAMP Reaction] Prepare the qPCR instrument, have Ice on hand to prepare the master Assay mix. Calculate the amount of samples b...
["[Sample Purification / Concentration]\nPreheat the Heat Block or Water Bath", "[RT-LAMP Reaction]\nPrepare the qPCR instrument, have Ice on hand to prepare the master Assay mix. Calculate the amount of samples being run and make sure to include the Positive Control and the H20.", "[Sample Lysis / Inactivation]\nCent...
59,054
2D TEM CLEM (Correlative Light Microscopy and Electron Microscopy)
4
dx.doi.org/10.17504/protocols.io.261gend2jg47/v1
https://www.protocols.io/view/2d-tem-clem-correlative-light-microscopy-and-elect-b5wnq7de
Pietro De Camilli, Yumei Wu
TITLE: 2D TEM CLEM (Correlative Light Microscopy and Electron Microscopy) AUTHORS: Pietro De Camilli, Yumei Wu [DESCRIPTION] This protocol details the general procedure of Correlative Light Microscopy and Electron Microscopy (CLEM) with conventional chemical fixation and 2D Transmission Electron Microscopy (TEM) imagi...
["[General preparation:] Culture cells on poly-d-lysine-coated 35 mm MatTek dish (P35G-1.5-14-CGRD) and transfect the cells with the plasmids of the interest.", "[General preparation:] Pre-fix the cells in 37 °C-warmed 4% PFA and 0.25% glutaraldehyde in the imaging buffer.", "[General preparation:] Wash in the same ima...
94,272
SOP for Flow cytometry after DSS-induced gut and brain injury
4
dx.doi.org/10.17504/protocols.io.bp2l6x6zklqe/v1
https://www.protocols.io/view/sop-for-flow-cytometry-after-dss-induced-gut-and-b-c8a8zshw
Malu G Tansey
TITLE: SOP for Flow cytometry after DSS-induced gut and brain injury AUTHORS: Malu G Tansey [DESCRIPTION] SOP for Flow cytometry after DSS-induced gut and brain injury [STEPS] 1. Following isolation of brain and circulating immune cells with magnetic beads, cells were transferred to a V-bottom plate for surface epito...
["Following isolation of brain and circulating immune cells with magnetic beads, cells were transferred to a V-bottom plate for surface epitope labeling.", "Samples were centrifuged at 300g for 5min at 4°C, washed once in PBS, re-pelleted, and resuspended in an antibody cocktail containing fluorophore-conjugated antibo...
null
null
null
dx.doi.org/10.17504/protocols.io.jcdcis6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This is an effective protocol for genotyping diatom exconjugants which we believe is readily applicable to any other genetically manipulated marine microeukaryote.</p> [BEFORE_START] <p>1. Your exconjugant colonies should be growing as small - ~300 µL - liquid cultures in th...
[]
68,430
Secondary Data Analysis - Creating a MycoMap Project from ONT Amplicon/Barcode Data
5
null
https://www.protocols.io/view/secondary-data-analysis-creating-a-mycomap-project-ce3ntgme
Stephen Douglas Russell
TITLE: Secondary Data Analysis - Creating a MycoMap Project from ONT Amplicon/Barcode Data AUTHORS: Stephen Douglas Russell [DESCRIPTION] This protocol outlines many of the steps that can be taken after the primary data analysis to expedite the final analysis - turning your data into results. Topics include c...
["[Create a MycoMap Project] Login to MycoMap.com. If you are not a member: \n\nRegister for a username on MycoMap.com: https://mycomap.com/login\n\nThen:\n\nCreate a project at the following link: https://mycomap.com/projects/create\n\nFill out the fields below:\n\nName: The name of your project. Something like \"Myco...
77,517
A single guide to impregnate samples with Golgi-Cox solution within 24hr and represent results with a set of algorithm
4
dx.doi.org/10.17504/protocols.io.3byl4jdrolo5/v1
https://www.protocols.io/view/a-single-guide-to-impregnate-samples-with-golgi-co-cpxmvpk6
Avishek Roy, Binney Sharma
TITLE: A single guide to impregnate samples with Golgi-Cox solution within 24hr and represent results with a set of algorithm AUTHORS: Avishek Roy, Binney Sharma [DESCRIPTION] Golgi-Cox staining is one of the old but relevant histological technique to identify neurons in superficial/ deep brain structures. The goal of...
["[Transcardial perfusion] Perfusion setup was filled with 0.9 % NaCl (saline) 2-4 °C and the flow rate was set at a rate of 3ml/min", "[Transcardial perfusion] Rats were placed onto their back and heart was made visible by opening cardiac envelop followed by an access to plural cavity through incisions through diaphr...
94,571
Preparation and RNAscope-labeling of fresh mouse midbrain tissue
1
dx.doi.org/10.17504/protocols.io.eq2lyjynelx9/v1
https://www.protocols.io/view/preparation-and-rnascope-labeling-of-fresh-mouse-m-c8kjzuun
William S Conrad
TITLE: Preparation and RNAscope-labeling of fresh mouse midbrain tissue AUTHORS: William S Conrad [DESCRIPTION] Below we describe preparing and labeling coronal sections of mouse substantia nigra (SNc) and ventral tegmental area (VTA) for probes targeting the genes Slc17a6 (VGLUT2), Slc32a1 (VGAT), and Slc18a2 (VMAT2)...
["[A.\tMouse brain extraction] Add approximately 50mL of isopentane in a beaker (cleaned with RNAseZap) and chill on dry ice.", "[A.\tMouse brain extraction] Anaesthetize mouse with pentobarbital (200 mg/kg i.p.).", "[A.\tMouse brain extraction] Decapitate mouse and extract brain from skull.", "[A.\tMouse brain extract...
18,686
Nutrient solution for rice hydroponics culture
1
dx.doi.org/10.17504/protocols.io.wg6fbze
https://www.protocols.io/view/nutrient-solution-for-rice-hydroponics-culture-wg6fbze
John Platten
TITLE: Nutrient solution for rice hydroponics culture AUTHORS: John Platten [DESCRIPTION] This represents a heavily modified Yoshida's solution. Compared to Yoshida's formulation, this solution has almost the same concentration of macro and micronutrients, but the formulation has been simplified. Key advantages: * T...
["[Solution A, 1000× (Main macro and micronutrients)] Make this solution up as a SINGLE stock solution. For 1L of stock solution,\n* Start with ~600mL de-ionised water.\n* Weigh out and dissolve each component (except ferric chloride and citric acid) directly in the 600mL solution. Allow each to dissolve completely b...
null
null
null
dx.doi.org/10.17504/protocols.io.unpevdn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Caenorhabditis elegans, a small (adults are ~1 mm long), free-living soil nematode that feeds on bacteria, is an ideal organism for applying various live microscopy techniques. The organism is transparent, thus it is possible to microscopically analyze and record the whole anima...
["[Pre-Imaging set-up] Clean the rectangular glass plate with ethanol and lint free tissue\n Switch ON the rig \n Align the movable part till desired position and lock it by pressing ON (If you want to move it again press EMO, to lock again twist the EMO knob and press ON again)\n Log in to the 3 PCs (Password: BehavGe...
71,945
Solid fungal extraction and C8 reversed phase chromatography
6
dx.doi.org/10.17504/protocols.io.ewov1onpklr2/v1
https://www.protocols.io/view/solid-fungal-extraction-and-c8-reversed-phase-chro-cihhub36
Shara Van De Pas, Siouxsie Wiles
TITLE: Solid fungal extraction and C8 reversed phase chromatography AUTHORS: Shara Van De Pas, Siouxsie Wiles [DESCRIPTION] Protocol for freeze-drying fungal cultures and preparing extracts using C8 reversed-phase chromatography. [STEPS] SECTION: Crude extraction of fungal compounds 1. Subculture fungus onto ~40 Petr...
["[Crude extraction of fungal compounds] Subculture fungus onto ~40 Petri dishes of media of choice and seal the plates with parafilm. Grow at the appropriate growth temperature until the fungus reaches the required age or coverage.", "[Crude extraction of fungal compounds] Weigh a clean 500ml beaker and break up dry f...
66,016
Functionality test (DNA gel stain)
4
null
https://www.protocols.io/view/functionality-test-dna-gel-stain-ccp8svrw
Nadine Mowoh, Stephane Fadanka
TITLE: Functionality test (DNA gel stain) AUTHORS: Nadine Mowoh, Stephane Fadanka [DESCRIPTION] After PCR amplification of DNA, agarose gel electrophoresis is run to separate the DNA based on their size. The agarose gel consists of microscopic pores that act as a molecular sieve which separates molecules based on t...
["[Functionality test of Thiazole Orange-DMSO based DNA gel stain] Preparing 1.5 % agarose gel (using the test DNA gel stain)\n\nPrepare a 1.5 % agarose gel for electrophoresis as follows:\n\n \n\n \n\n\nUse a weighing balance to weigh 0.375 gof agarose powder and pour into a 150 ml or any appropriate size glass beaker...
26,565
A rapid, sensitive, scalable method for Precision Run-On sequencing (qPRO-seq)
null
dx.doi.org/10.17504/protocols.io.57dg9i6
https://www.protocols.io/view/a-rapid-sensitive-scalable-method-for-precision-ru-57dg9i6
Julius Judd, Luke A. Wojenski, Lauren M. Wainman, Nathaniel D. Tippens, Edward J. Rice, Alexis Dziubek, Geno J. Villafano, Erin M. Wissink, Philip Versluis, Lina Bagepalli, Sagar R. Shah, Dig B. Mahat, Jacob M. Tome, Charles G. Danko, John T. Lis, Leighton J. Core
TITLE: A rapid, sensitive, scalable method for Precision Run-On sequencing (qPRO-seq) AUTHORS: Julius Judd, Luke A. Wojenski, Lauren M. Wainman, Nathaniel D. Tippens, Edward J. Rice, Alexis Dziubek, Geno J. Villafano, Erin M. Wissink, Philip Versluis, Lina Bagepalli, Sagar R. Shah, Dig B. Mahat, Jacob M. Tome, Charles ...
["[Cell Permeabilization]\nPrepare permeabilization buffer, wash buffer, and freeze buffer and place .\non ice\nCAUTION: DEPC is toxic and harmful!\nCRITICAL: Care should be taken to avoid nuclease contamination. Change gloves routinely and prepare/use nuclease-free reagents.\nCRITICAL: ALL steps should be carried out ...
63,730
Diabetic Code Protocol
5
dx.doi.org/10.17504/protocols.io.261genj8wg47/v1
https://www.protocols.io/view/diabetic-code-protocol-cagssbwe
Carlos A Maldonado Virella
TITLE: Diabetic Code Protocol AUTHORS: Carlos A Maldonado Virella [DESCRIPTION] installs of the cure [STEPS] 1. 2. 3. 4.
[]
87,926
Soil extract for cocultures host-parasites media
1
dx.doi.org/10.17504/protocols.io.rm7vzxkq2gx1/v1
https://www.protocols.io/view/soil-extract-for-cocultures-host-parasites-media-cz4wx8xe
Roscoff Culture Collection, Estelle Bigeard, Laure Guillou
TITLE: Soil extract for cocultures host-parasites media AUTHORS: Roscoff Culture Collection, Estelle Bigeard, Laure Guillou [DESCRIPTION] How to prepare soil extract [BEFORE_START] Please refer to our general recommendations to grow cultures : https://www.protocols.io/private/A48906DC1374AD6281495CB86A8F092F [STE...
["[Preparation of the soil extract solution (25g/l)] Add 1200ml of MilliQ water to the soil. \nClose the erlenmeyer with a aluminium foil.", "[Preparation of the soil extract solution (25g/l)] Heat at 100°C with pemanent mixing during 1 hour to obtain an efficient boiling.\nAdjust this time depending on the volume (mor...
null
null
null
dx.doi.org/10.17504/protocols.io.jh7cj9n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Set up ssh keys to make login to the HPC easy!</p> [STEPS] ?. ?. ?. ?. ?. ?.
[]
101,280
Use of EcoFAB 2.0 for reproducible plant-microbe interaction experiments
0
dx.doi.org/10.17504/protocols.io.kxygxyydkl8j/v1
https://www.protocols.io/view/use-of-ecofab-2-0-for-reproducible-plant-microbe-i-de583g9w
Vlastimil Novak, Eoghan King, Peter Andeer, John Vogel, Trent t Northen
TITLE: Use of EcoFAB 2.0 for reproducible plant-microbe interaction experiments AUTHORS: Vlastimil Novak, Eoghan King, Peter Andeer, John Vogel, Trent t Northen [DESCRIPTION] We conducted a multi-laboratory experiment across five labs to demonstrate reproducible microbiome formation, metabolite production, and plant g...
["[Materials and equipment] Materials sent by the organizing lab (LBNL)\nMaterial needed for 40 EcoFAB (See EcoFAB 2.0 assembly protocol)\nSterilization pouches\nMurashige & Skoog (MS) Basal Salts (Caisson Labs MSP01-1LT).\nMicropore tape (UV-treated)\n15 mL centrifuge tubes, sterile\n2 mL microcentrifuge tubes contain...
28,925
Protocol for Subculture of Differentiated Blood-Brain Barrier Endothelial Cells onto Plates and Filters
null
dx.doi.org/10.17504/protocols.io.8g5hty6
null
Ethan Lippmann, Hannah Wilson, Emma Neal
TITLE: Protocol for Subculture of Differentiated Blood-Brain Barrier Endothelial Cells onto Plates and Filters AUTHORS: Ethan Lippmann, Hannah Wilson, Emma Neal [STEPS] ?. [Subculturing (Day 6)] Please select between subculturing onto plates or filters. ?. [Subculturing (Day 6)] Coat plates with ECM plate solution for...
["[Subculturing (Day 6)]\nPlease select between subculturing onto plates or filters.", "[Subculturing (Day 6)]\nCoat plates with ECM plate solution for at least at . Volume depends on plate type (see Table): ABC1Plate type for subculture phaseVolume of ECM solution for coatingWorking volume of EC media for cell cultu...
68,472
Transforming E. coli
4
null
https://www.protocols.io/view/transforming-e-coli-ce4ytgxw
Brian Teague
TITLE: Transforming E. coli AUTHORS: Brian Teague [DESCRIPTION] Transformation is the process of inducing chemically competent E. coli to take up a plasmid. We do this in order to use E. coli as a "DNA copier" -- the bacterium takes up the plasmid, then copies it as it copies its own genomic DNA. We can grow up as ...
["[Transformation] Retrieve the two tubes of chemically competent E. coli cells from the -80 °C freezer and place them immediately on ice. Incubate the cells on ice 3-4 minutes to thaw.", "[Transformation] Add 1 µL of DNA from your ligation to a tube of cells. Immediately after adding the DNA to the tube, flick several...
40,318
Preparation of a protein-AG conjugated to horseradish peroxidase by the periodate method.
6
dx.doi.org/10.17504/protocols.io.bjk6kkze
https://www.protocols.io/view/preparation-of-a-protein-ag-conjugated-to-horsera-bjk6kkze
Angel Justiz-Vaillant
TITLE: Preparation of a protein-AG conjugated to horseradish peroxidase by the periodate method. AUTHORS: Angel Justiz-Vaillant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A recombinant protein that combines the IgG-binding domains of SpA and SpG was developed, and labelled to horseradish pero...
["Horseradish peroxidase (500 µg in 50 µl NaCO3 , pH 9.6) is mixed with freshly made sodium periodate solution (1.71 mg/ml) followed by incubation in the dark for 2 h.", "Mix 500 µg of staphylococcal protein-A (SpA) with an equal amount (500 micrograms) of a mix of horseradish peroxidase-sodium periodate. On the other ...
44,037
Vivarium Population Spender: Immigration Module
5
dx.doi.org/10.17504/protocols.io.bn9dmh26
https://www.protocols.io/view/vivarium-population-spender-immigration-module-bn9dmh26
Camila Rangel Smith
TITLE: Vivarium Population Spender: Immigration Module AUTHORS: Camila Rangel Smith [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Description of the steps followed by Vivarium Population Spenser library when running the Immigration module. </div></div> [STEPS] ?. Get the total number of immigrant...
["Get the total number of immigrants expected for that local authority on a year from a local authority level international migration table.", "Divide the number of total immigrants to be assigned by the number of steps existing in that year.", "For each time step:", "Add the fraction of immigrants to be assigned in on...
53,999
Glycerol stock preparation
4
dx.doi.org/10.17504/protocols.io.byyppxvn
https://www.protocols.io/view/glycerol-stock-preparation-byyppxvn
Ashwinuday
TITLE: Glycerol stock preparation AUTHORS: Ashwinuday [DESCRIPTION] This protocol is for the preparation of 10% glycerol stocks of bacterial strains. [STEPS] 1. From a streaked plate of the bacteria, pick one colony and inoculate into a test tube containing 10 mL 1X LB 2. Grow the culture till it reaches an OD ~ ...
["From a streaked plate of the bacteria, pick one colony and inoculate into a test tube containing 10 mL 1X LB", "Grow the culture till it reaches an OD ~ 0.6", "Into cryo-vials add 1.6 mL culture and 0.4 mL 50% autoclaved glycerol.", "Flash freeze the tubes in liquid Nitrogen and store it at -80 °C"]
76,285
TaME-seq2
1
dx.doi.org/10.17504/protocols.io.dm6gpjxy5gzp/v1
https://www.protocols.io/view/tame-seq2-cnq5vdy6
jean-marc, Milan Stosic, Alexander Hesselberg, trinro
TITLE: TaME-seq2 AUTHORS: jean-marc, Milan Stosic, Alexander Hesselberg, trinro [DESCRIPTION] Tagmentation-assisted multiplex PCR enrichment sequencing for viral genomic profiling (TaME-seq2). Viral genome and integration enrichment library preparation protocol. Manuscript: TaME-seq2: Tagmentation-assisted multiplex...
["[Sample preparation] Prepare and normalize DNA/cDNA samples by measuring sample concentration and diluting in nuclease-free water if necessary. \n\nSample volume should be 15 µL \n\n50 ng input is recommended, but the protocol works with less and performance is more dependent on viral load.", "[Tagmentation] Prepare...
100,178
Bacterial genome annotation script using BLASTN
5
dx.doi.org/10.17504/protocols.io.dm6gpjrb1gzp/v3
https://www.protocols.io/view/bacterial-genome-annotation-script-using-blastn-dd3s28ne
Ana Mariya Anhel, Lorea Alejaldre, Ángel Goñi-Moreno
TITLE: Bacterial genome annotation script using BLASTN AUTHORS: Ana Mariya Anhel, Lorea Alejaldre, Ángel Goñi-Moreno [DESCRIPTION] This protocol uses a python based script and command-line BLASTn to annotate in a final table single-read sequencing results from genome amplifications, within other output files. Its mai...
["[Adquisition of files] Download reference genome file\n\nYou can download the genome of the organism that you want to compare to the reading sequences from different sources such as NCBI, GSA or even pages dedicated to the organism (for example pseudomonas.com).\n\nFor this script to work, genome files need to be in ...
22,116
Squalene Quantification using Nile Red Staining (Developmental)
null
dx.doi.org/10.17504/protocols.io.zucf6sw
null
Sebastian Triesch
TITLE: Squalene Quantification using Nile Red Staining (Developmental) AUTHORS: Sebastian Triesch [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">This protocol is under development!</span></div><div class = "text-block">Nile Red is a fluorescent dye that stains sele...
["[Culture Sampling]\nSample 1-2 ml Synechocystis culture, measure its OD at 750 nm and adjust it to 2 ml of OD (750 nm) = 0.5 in BG-11 media. Split your adjusted culture in 2x 1 ml. One portion will be stained with Nile Red, the other will serve as a negative control.", "[Staining]\nStain one portion of the previously...
71,353
Constant mood monitoring using WhatsApp for persons with depression: a pilot study
2
dx.doi.org/10.17504/protocols.io.8epv5jz46l1b/v1
https://www.protocols.io/view/constant-mood-monitoring-using-whatsapp-for-person-chwzt7f6
Maritta A Välimäki, Qing Long, Min Yang, Sau Fong
TITLE: Constant mood monitoring using WhatsApp for persons with depression: a pilot study AUTHORS: Maritta A Välimäki, Qing Long, Min Yang, Sau Fong [DESCRIPTION] Background Preliminary evidence suggests that social media could provide a convenient platform to monitor mood. It is still unknown whether WhatsApp mood mo...
[]
81,176
Simple TCA/acetone protein extraction protocol for proteomics studies.
4
dx.doi.org/10.17504/protocols.io.5jyl8j74dg2w/v1
https://www.protocols.io/view/simple-tca-acetone-protein-extraction-protocol-for-cthywj7w
Angelo José Rinaldi
TITLE: Simple TCA/acetone protein extraction protocol for proteomics studies. AUTHORS: Angelo José Rinaldi [DESCRIPTION] Protein extraction with TCA (trichloroacetic acid) and acetone is a widely used method to precipitate proteins from biological samples. This method is useful for obtaining proteins of interest for p...
["Prepare a 20% TCA solution by adding 200g of TCA in 800ml of distilled water. Be sure to do this in a well-ventilated area, wearing gloves and eye protection, as TCA is corrosive.", "Add the biological sample (approximately 300 mg) to a 2 mL microcentrifuge tube.", "Add a small amount of detergent (10% SDS) to the sa...
28,611
Removal of gDNA out of totalRNA
null
dx.doi.org/10.17504/protocols.io.77bhrin
null
iGEM Dusseldorf
TITLE: Removal of gDNA out of totalRNA AUTHORS: iGEM Dusseldorf [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Start</span></div><div class = "text-block">Calculate the volume required for 1 µg RNA</div><div class = "text-block"><span style = "font-weight:bold;">Di...
[]
60,846
H&amp;E Staining for Pancreas or Eye Cryosections
1
dx.doi.org/10.17504/protocols.io.rm7vz3e6rgx1/v2
https://www.protocols.io/view/h-amp-e-staining-for-pancreas-or-eye-cryosections-b7nnrmde
Diane Saunders, Angela Kruse, Jamie Allen, Carrie Romer, Danielle Gutierrez, Alvin Powers, Jeff Spraggins
TITLE: H&amp;E Staining for Pancreas or Eye Cryosections AUTHORS: Diane Saunders, Angela Kruse, Jamie Allen, Carrie Romer, Danielle Gutierrez, Alvin Powers, Jeff Spraggins [DESCRIPTION] This protocol for H&E staining can be applied to either fixed or unfixed frozen cryosections. [STEPS] 1. Air dry sections for...
["Air dry sections for 5 min .", "Incubate in 95% alcohol for 1 min , then wash, dipping until clear.", "Incubate in hematoxylin for 30 s , then wash until clear.", "Dip 3-4x in bluing solution, then wash for 30 s", "Dip 3-4x in 95% alcohol.", "Dip 1-2x in eosin.", "Dip 5-10x in 95% alcohol, then repeat using 2nd aliqu...
33,152
PROTOCOL FOR: LRP37 method
null
dx.doi.org/10.17504/protocols.io.bck8iuzw
null
Gaku Takahashi, Katsuya Inada
TITLE: PROTOCOL FOR: LRP37 method AUTHORS: Gaku Takahashi, Katsuya Inada [STEPS]
[]
101,699
CYToF Staining
0
dx.doi.org/10.17504/protocols.io.14egn69nml5d/v1
https://www.protocols.io/view/cytof-staining-dfjb3kin
Meelad Amouzgar, Patricia Favaro, Daniel Ho, Trevor Bruce, Kausalia Vijayaragavan, Sean Bendall
TITLE: CYToF Staining AUTHORS: Meelad Amouzgar, Patricia Favaro, Daniel Ho, Trevor Bruce, Kausalia Vijayaragavan, Sean Bendall [DESCRIPTION] CYToF has significantly advanced immunophenotyping, especially in exploratory settings where comprehensive characterization of immune populations is necessary but sample sizes ar...
["[CyToF Staining] PFA Fixation:\nAdd 100 uL of 16% PFA per 1 mL of media and incubate cells for 10 minutes at room temperature.\nWash with 4mL of CSM 2x\nPellet the cells by centrifugation at 600xg for 5 min.\nStore fixed cells in 1mL of CSM at -80C", "[CyToF Staining] Barcoding protocol:\nTransfer cells to cluster t...
18,474
N. parisii infection of C. elegans
null
dx.doi.org/10.17504/protocols.io.waiface
null
Emily Troemel
TITLE: N. parisii infection of C. elegans AUTHORS: Emily Troemel [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.frsbm6e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The Sequence Read Archive (SRA) is a database for biological sequence data and is maintained by the National Center for Biotechnology Information (NCBI). Sequence files can be obtained by using the SRA Toolkit. This protocol provides the steps necessary to use the SRA toolkit...
[]
24,576
Immunofluorescence Staining of Sea Urchin Embryos
null
dx.doi.org/10.17504/protocols.io.388grzw
null
David Booth
TITLE: Immunofluorescence Staining of Sea Urchin Embryos AUTHORS: David Booth [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">In this lab, you will use immunofluroescence stainging to visualize the wondrous cellular transformations that occur throughout sea urchin development. The goals of this modu...
["[Fix Cells]\nPrepare cells for fixation• Place 300 µl of embryos in filtered seawater in 1 well of a 96-well, round-bottom dish.", "[Add Primary Antibody]\nPrepare primary antibody• Dilute mouse anti-Tubulin antibody 1:500 in Block solution! Remember that you will need 200 µl of primary antibody per well", "[Block]\n...
62,194
Cholera Toxin Subunit B (CTB) Retrograde tracing from the mouse colon and bladder wall.
1
dx.doi.org/10.17504/protocols.io.x54v9y391g3e/v1
https://www.protocols.io/view/cholera-toxin-subunit-b-ctb-retrograde-tracing-fro-b8ysrxwe
Andrea Harrington
TITLE: Cholera Toxin Subunit B (CTB) Retrograde tracing from the mouse colon and bladder wall. AUTHORS: Andrea Harrington [DESCRIPTION] This protocol is used to label the neurons and their central terminals of the sensory neurons innervating the colon and bladder in an experimental adult male or female mouse. The pr...
["[Preparation for surgery] Prepare cages for each mouse – place clean bedding, nesting material in cage and water bottle and food pellets in hopper ready for post-surgery", "[Anaesthesia] Place mouse in induction box, and turn on isoflurane to 5% and oxygen flow rate to 0.5-0.6L/min.", "[Preparation for surgery] Weigh...
54,952
DNA extraction from concrete
1
null
https://www.protocols.io/view/dna-extraction-from-concrete-bzwgp7bw
Anders Kiledal, Julia A Maresca
TITLE: DNA extraction from concrete AUTHORS: Anders Kiledal, Julia A Maresca [DESCRIPTION] This is a protocol for extracting DNA from concrete, based on the protocol developed by L. S. Weyrich, et al. for extraction of DNA from ancient calcified dental plaque. We have scaled it up for larger sample sizes and made som...
["[Sample Pre-Treatment (Day 1)] In a 50 mL conical tube, place 10 g pulverized concrete, 5 mL 0.5 M EDTA, 150 µL Proteinase K (20 mg/mL), 138 µL 20% SDS, and 0.2 mL glacial acetic acid. Incubate at 55°C overnight with gentle rotation.", "[DNA Extraction (Days 2 and 3)] Vortex at maximum speed on a platform vortexer fo...
null
null
null
dx.doi.org/10.17504/protocols.io.utcewiw
null
null
TITLE: No Title AUTHORS: [BEFORE_START] Thaw GeltrexTM Basement Membrane Matrix at 2-8°C overnight. Refrigerator temperatures may vary; therefore, thaw extract on ice in a refrigerator. Basement Membrane Matrix gels in 5-10 minutes above 15°C; therefore when working from a full 5 ml vial, it is unnecessary to keep it...
["[Thawing of Geltrex, dilution in DMEM/F12 and aliquotting for storage at -20C] {\"blocks\":[{\"key\":\"cvdtr\",\"text\":\" \",\"type\":\"unstyled\",\"depth\":0,\"inlineStyleRanges\":[],\"entityRanges\":[{\"offset\":0,\"length\":1,\"key\":0}],\"data\":[]},{\"key\":\"9u3ma\",\"text\":\"Thaw one tube of Geltrex (1-5 mL)...
23,758
Determination of Glomerular Filtration Rate in Conscious Mice using FITC-inulin
null
dx.doi.org/10.17504/protocols.io.3fngjme
null
Zhonghua Qi, Matthew D. Breyer
TITLE: Determination of Glomerular Filtration Rate in Conscious Mice using FITC-inulin AUTHORS: Zhonghua Qi, Matthew D. Breyer [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This protocol describes the procedures for ...
["Preparation of 5% FITC-inulin solution:1. 5% FITC-inulin is dissolved in two ml of 0.9% NaCl -- facilitated by heating the solution in boiling water. 2. To remove residual FITC not bound to inulin, the solution is filled into a 1000 Daltons cut-off dialysis membrane (Spectra/Pro 6, Spectrum Laboratories Inc., Rancho ...
null
null
null
dx.doi.org/10.17504/protocols.io.fyzbpx6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div class="page" title="Page 1"> <div class="layoutArea"> <div class="column"> <p>The SensiFAST™ SYBR® &amp; Fluorescein Kit uses a combination of the latest advances in buffer chemistry and enhancers, together with an antibody-mediated hot-start DNA polymerase system, to ensur...
[]
78,808
Compartmental Protein Extraction to Achieve Enrichment of Extracellular Matrix (ECM) Proteins
4
dx.doi.org/10.17504/protocols.io.kxygx94b4g8j/v1
https://www.protocols.io/view/compartmental-protein-extraction-to-achieve-enrich-cq7yvzpw
James Considine, Ikram Isa, Alexandra Naba
TITLE: Compartmental Protein Extraction to Achieve Enrichment of Extracellular Matrix (ECM) Proteins AUTHORS: James Considine, Ikram Isa, Alexandra Naba [DESCRIPTION] This protocol provides instructions on how to extract cellular components of tissues and enrich for ECM proteins to be later analyzed via immunoblotting...
["[Tissue Homogenization] Homogenize 100 mg of tissue in 1 mL of Buffer CEB containing protease inhibitors using a tissue homogenizer until the tissue is completely disrupted and a homogenous suspension is obtained.", "[Sequential extraction of intracellular soluble proteins]", "[Sequential extraction of intracellula...
23,884
Neuropathy Phentoyping Protocols - Apoptag (Peroxidase) Sections on Slides
null
dx.doi.org/10.17504/protocols.io.3jkgkkw
null
Eva Feldman
TITLE: Neuropathy Phentoyping Protocols - Apoptag (Peroxidase) Sections on Slides AUTHORS: Eva Feldman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block"><span style = "font-weight:bold;">Phenotyping of Rodents for the Pre...
["[Preparation:]\n1. Thaw equilibration buffer, TdT, reaction buffer and stopwash on ice. 2. Prepare working strength TdT enzyme. 3. Thaw enough reaction buffer and TdT to cover 30 ul/well TdT: reaction buffer, 33 µl: 77 µl. 4. Prepare stopwash buffer, add 1 ml buffer to 34 ml of ddH²0. 5. Prepare a humidified chamber....
null
null
null
dx.doi.org/10.17504/protocols.io.te7ejhn
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
99,977
Intraperitoneal Injection in an Adult Mouse
1
dx.doi.org/10.17504/protocols.io.5qpvo5w8dl4o/v2
https://www.protocols.io/view/intraperitoneal-injection-in-an-adult-mouse-ddvh2636
Allen Institute for Brain Science
TITLE: Intraperitoneal Injection in an Adult Mouse AUTHORS: Allen Institute for Brain Science [DESCRIPTION] This protocol describes the general procedure used for intraperitoneal injection in adult mice. Note: Research reported in this publication was supported by the National Institute Of Mental Health of the Nation...
[]
82,822
An axenic plant culture system for Sporobolus alterniflorus
4
dx.doi.org/10.17504/protocols.io.x54v9d94qg3e/v1
https://www.protocols.click/view/an-axenic-plant-culture-system-for-sporobolus-alte-cu5ewy3e
Elena L. Peredo, Suzanne M Thomas, Zoe Cardon
TITLE: An axenic plant culture system for Sporobolus alterniflorus AUTHORS: Elena L. Peredo, Suzanne M Thomas, Zoe Cardon [DESCRIPTION] Sporobulus alterniflorus is a native grass that dominates intertidal salt marsh platforms along thousands of miles of the U.S. East and Gulf coasts. This grass exhibits tolerance to ...
["[Seed storage and germination] The plant material in this protocol is seedlings germinated ex vitro. While optional, we recommend autoclaving the soil and completing surface sterilization of the seeds to minimize the contaminants in the samples. Alternatively, seeds can be germinated in open containers.", "[Seed stor...
74,205
Nano3P-seq Protocol
4
dx.doi.org/10.17504/protocols.io.3byl4j292lo5/v2
https://www.protocols.io/view/nano3p-seq-protocol-ckp5uvq6
Oguzhan Begik
TITLE: Nano3P-seq Protocol AUTHORS: Oguzhan Begik [DESCRIPTION] Here, we develop Nanopore 3’ end-capture sequencing (Nano3P-seq), a novel method that relies on nanopore cDNA sequencing to simultaneously quantify RNA abundance, tail composition and tail length dynamics at per-read resolution. By employing a template sw...
["[Without Barcoding] Materials and consumables required\n\n●Direct cDNA Sequencing Kit (ONT, SQK-DCS109)\n●Flow Cell Priming Kit (ONT, EXP-FLP001)\n●AMPure XP Reagent (Agencourt , A63881)\n●Blunt/TA Ligase Master Mix (NEB, M0367)\n●TGIRT™-III Enzyme (InGex)\n●RNase Inhibitor, Murine (NEB,M0314L)\n●RNase Cocktail Enzy...
91,049
Ex vivo mouse brain patch clamp recordings combined with uncaging and optogenetic stimulation
4
dx.doi.org/10.17504/protocols.io.rm7vzx1w2gx1/v2
https://www.protocols.io/view/ex-vivo-mouse-brain-patch-clamp-recordings-combine-c46hyzb6
enrico.zampese
TITLE: Ex vivo mouse brain patch clamp recordings combined with uncaging and optogenetic stimulation AUTHORS: enrico.zampese [DESCRIPTION] In this protocol we detail the steps to perform ex-vivo brain slices electrophysiology and optogenetic stimulation and/or 2-photon uncaging recordings. [BEFORE_START] Please note ...
["[Prepare patch pipettes] Turn on the Sutter P-1000 puller and enter the desired pull protocol.", "[Prepare patch pipettes] Insert a thick-walled borosilicate glass capillary and press pull.", "[Prepare patch pipettes] Pipette resistance must be of 3 to 5 megaohms.", "[Setting up patch rig and environment] Turn on the...
35,684
FACS Single Cell Sorting
null
dx.doi.org/10.17504/protocols.io.be4cjgsw
null
Allen Institute for Brain Science
TITLE: FACS Single Cell Sorting AUTHORS: Allen Institute for Brain Science [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol provides step by step instructions for performing single-cell sorting by using BD FACSAria II machine or BD FACSAria Fusion machine and BD FACSDiva software.</div>...
[]
27,208
Toxicity assay for inducer compounds in Synechocystis sp. PCC 6803
null
dx.doi.org/10.17504/protocols.io.6tghejw
null
Anna Behle, Pia Saake, Ilka Maria Axmann
TITLE: Toxicity assay for inducer compounds in Synechocystis sp. PCC 6803 AUTHORS: Anna Behle, Pia Saake, Ilka Maria Axmann [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Inducible promoters are an important tool for synthetic biology. They enable temporal control of gene expression, as well as con...
["[Inoculation of pre-culture]\nInoculate an appropriate amount of BG11 with your cyanobacterial strain of choice. The inoculation volume depends on the number of concentrations to be tested. For example: 3 concentrations x 3 replicates x 30 mL = 270 mL of preculture. Since the culture will be diluted ~1:3 before start...
44,614
Automated high throughput viral concentration from wastewater using the KingFisher Flex platform
4
null
https://www.protocols.io/view/automated-high-throughput-viral-concentration-from-bptemnje
Smruthi Karthikeyan, Greg Humphrey
TITLE: Automated high throughput viral concentration from wastewater using the KingFisher Flex platform AUTHORS: Smruthi Karthikeyan, Greg Humphrey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Large-scale wastewater surveillance has the ability to greatly augment the tracking of infection dynami...
["Aliquot of raw sewage (no pre-filtering step required) into two wells of the KingFisher 24 deep-well plate (Cat# 95040470, Thermo Fisher). Each well should have of the raw sewage sample. For instance, of sample PL09 should be filled in A1 of plate 1 and in A1 of plate 2.\n10 mL\n5 mL\n5 mL\n5 mL", "Add of t...
54,436
Step 1: Swab sample collection
1
null
https://www.protocols.io/view/step-1-swab-sample-collection-bzecp3aw
Peter H L Krijger, Tim A Hoek, Sanne Boersma, Lieke I P M Donders, Maaike M C Broeders, Mark Pieterse, Pim W Toonen, Ive Logister, Bram M P Verhagen, Marjon J A M Verstegen, Thomas W van Ravesteyn, Rene J T M Roymans, Francesca Mattiroli, Jo Vandesompele, Monique Nijhuis, Stefan Meijer, Anton van Weert, Edwin Dekker, F...
TITLE: Step 1: Swab sample collection AUTHORS: Peter H L Krijger, Tim A Hoek, Sanne Boersma, Lieke I P M Donders, Maaike M C Broeders, Mark Pieterse, Pim W Toonen, Ive Logister, Bram M P Verhagen, Marjon J A M Verstegen, Thomas W van Ravesteyn, Rene J T M Roymans, Francesca Mattiroli, Jo Vandesompele, Monique Nijhuis, ...
["[Sample collection] Person 1 scans the 2D barcode at the bottom of a 0.8 ml Micronic tube prefilled with 0.4 ml virus transport buffer ( InActiv Blue mixed with a spike-in control RNA) and places it in a Micronic tube rack (tube rack A).", "[Sample collection] Person 1 removes cap from the Micronic tube using the man...
83,898
NEBNext Ultra II FS DNA Module E7810
1
dx.doi.org/10.17504/protocols.io.bp2l66xrlqe5/v2
https://www.protocols.click/view/nebnext-ultra-ii-fs-dna-module-e7810-cv62w9ge
jbonnevie
TITLE: NEBNext Ultra II FS DNA Module E7810 AUTHORS: jbonnevie [DESCRIPTION] The NEBNext Ultra II FS DNA Module contains the enzymes and buffers required to convert a broad range of input amounts of intact DNA into fragmented DNA with 5´ phosphorylated 3´ dA-tailed ends. The module is optimized for use with the NEBNex...
["[Fragmentation/End Prep] Fragmentation occurs during the 37 °C incubation step. Use the chart below to determine the incubation time required to generate the desired fragment sizes. Incubation time may need to be optimized for individual samples. See Figure 1 for a typical fragmentation pattern.\n \n Fragmentation ...
83,014
Astrocyte isolation and culturing
4
dx.doi.org/10.17504/protocols.io.e6nvwdp9wlmk/v1
https://www.protocols.io/view/astrocyte-isolation-and-culturing-cvbew2je
Julia F Riley
TITLE: Astrocyte isolation and culturing AUTHORS: Julia F Riley [DESCRIPTION] This protocol details steps for preparing an astrocyte monoculture from mouse pups. From dissection to plated samples ready to be transfected, treated, lysed, or imaged, the protocol takes 20-25 days. [BEFORE_START] The night before (could...
["[Dissection] Sacrifice mouse pups postnatal day (P) 1-3 via isoflourane overdose.", "[Dissection] Decapitate pups and transfer heads to a dish with 100 % (v/v) EtOH. Swirl to clean.", "[Dissection] Transfer heads to a new 10cm dish and place whole dish in a cell culture dish with a wider diameter. The surrounding dis...
57,738
Seeding nucleofected hPSCs in 96-well plates using limited dilution
4
dx.doi.org/10.17504/protocols.io.b4miqu4e
https://www.protocols.io/view/seeding-nucleofected-hpscs-in-96-well-plates-using-b4miqu4e
Hanqin Li, Yogendra Verma, Dirk Hockemeyer, Frank Soldner
TITLE: Seeding nucleofected hPSCs in 96-well plates using limited dilution AUTHORS: Hanqin Li, Yogendra Verma, Dirk Hockemeyer, Frank Soldner [DESCRIPTION] This protocol describes a standard procedure for seeding nucleofected human pluripotent stem cells in 96-well plates using limited dilution. This protocol follows...
["For an editing with 10% efficiency, prepare two 96-well MEFs plates with a density at 2 million cells/plate at least 1 day ahead. Do not shake the plate after seeding MEFs. It generates swirls and causes cells to accumulate at the center. If editing efficiency is expected to be low, prepare more 96-well plates.", "Ca...
49,085
Investigating DOIs&#39; classes of errors
5
dx.doi.org/10.17504/protocols.io.bt65nrg6
https://www.protocols.io/view/investigating-dois-39-classes-of-errors-bt65nrg6
Ricarda Boente, Deniz Tural, Cristian Santini, Arcangelo Massari
TITLE: Investigating DOIs&#39; classes of errors AUTHORS: Ricarda Boente, Deniz Tural, Cristian Santini, Arcangelo Massari [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The purpose of this protocol is to describe an automated process to repair invalid DOIs. In particular, four classes of errors ar...
["[Checking the DOI names' invalidity]\nWe import a CSV containing invalid DOI-to-DOI citations organized as follow: the first column contains valid citing DOIs and the second contains invalid cited DOIs", "[Checking the DOI names' invalidity]\nFirst, we check for each cited DOI if it is factually invalid. For this p...
27,980
Yeast transformation
null
dx.doi.org/10.17504/protocols.io.7jkhkkw
null
Marijn Ceelen
TITLE: Yeast transformation AUTHORS: Marijn Ceelen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a protocol for the transformation of yeast cells with linear or circular DNA.</div></div> [STEPS] ?. Make a transformation mix consisting of: AB1DNA34 µL (0.5-4 µg per fragment)2Salmon sperm ...
["Make a transformation mix consisting of: AB1DNA34 µL (0.5-4 µg per fragment)2Salmon sperm DNA 2 mg/mL50 µL3PEG-3350240 µL41M LiOAc36 µL5MQ waterup to a final volume of 360 µL\nAB1DNA34 µL (0.5-4 µg per fragment)2Salmon sperm DNA 2 mg/mL50 µL3PEG-3350240 µL41M LiOAc36 µL5MQ waterup to a final volume of 360 µL", "Cent...
null
null
null
dx.doi.org/10.17504/protocols.io.fcrbiv6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p> </p> <p><strong>Lysogeny <a href="https://en.wikipedia.org/wiki/Broth" target="_blank">broth</a></strong> (<strong>LB</strong>), a <a href="https://en.wikipedia.org/wiki/Nutrient" target="_blank">nutritionally</a> rich <a href="https://en.wikipedia.org/wiki/Growth_medium" ta...
[]
85,798
Gene editing of YIPF4 in hESCs V3
4
dx.doi.org/10.17504/protocols.io.rm7vzxy44gx1/v1
https://www.protocols.io/view/gene-editing-of-yipf4-in-hescs-v3-cx2exqbe
Harper JW, Kelsey Hickey, sharan_swarup
TITLE: Gene editing of YIPF4 in hESCs V3 AUTHORS: Harper JW, Kelsey Hickey, sharan_swarup [DESCRIPTION] This protocol describes the creation of YIPF4 knockout hESCs. [STEPS] SECTION: Cell line maintenance 1. Maintain H9 human embryonic stem cells (AAVS1-TRE3G-NGN2 hESCs) in complete (E8 media). SECTION: Targeted knoc...
["[Cell line maintenance] Maintain H9 human embryonic stem cells (AAVS1-TRE3G-NGN2 hESCs) in complete (E8 media).", "[Targeted knock-out of YIPF4] For YIPF4 knock-out, the following sgRNA sequences was designed and ordered from Synthego (5’ AAGAGGTTATGGCTGGCTTC 3').", "[Targeted knock-out of YIPF4] 0.6 μg sgRNA was in...
102,378
Behavioral Consequences of Dopaminergic Dysregulation Resulting from Violence-Induced Brain Trauma
0
dx.doi.org/10.17504/protocols.io.261ge5e17g47/v1
https://www.protocols.io/view/behavioral-consequences-of-dopaminergic-dysregulat-df8i3rue
Spencer Lee
TITLE: Behavioral Consequences of Dopaminergic Dysregulation Resulting from Violence-Induced Brain Trauma AUTHORS: Spencer Lee [DESCRIPTION] This study protocol outlines the experimental procedures designed to assess the influence of trauma resulting from violence on dopamine activation and subsequent behavior. The ex...
["[This study aimed to investigate dopaminergic dysregulation caused by trauma induced by violence, and its subsequent behaviors. This study will conduct a systematic review and meta-analysis and incorporate correlation statistical analysis.] The relationship between dopamine activation and subsequent dopamine release ...
22,996
Biolistic transformation of Isochrysis galbana
null
dx.doi.org/10.17504/protocols.io.2pugdnw
null
Glen Wheeler, Rowena Stern
TITLE: Biolistic transformation of Isochrysis galbana AUTHORS: Glen Wheeler, Rowena Stern [STEPS] ?. [Preparing tungsten beads] Weigh out 60 mg tungsten into a microfuge tube ?. [Preparing tungsten beads] Wash in 1 mL 100% ethanol. Vortex. Centrifugation at 13000rpm for 1 minute then remove ethanol ?. [Preparing tungs...
["[Preparing tungsten beads]\nWeigh out 60 mg tungsten into a microfuge tube", "[Preparing tungsten beads]\nWash in 1 mL 100% ethanol. Vortex. Centrifugation at 13000rpm for 1 minute then remove ethanol", "[Preparing tungsten beads]\nWash four times in 1 mL molecular grade water. Centrifugation at 13000rpm for 1 minute...
60,337
RT-qPCR Detection of Process Controls (Murine noroviurs and crAssphage) from Wastewater using AB 7500
4
dx.doi.org/10.17504/protocols.io.kqdg36j9pg25/v2
https://www.protocols.io/view/rt-qpcr-detection-of-process-controls-murine-norov-b66rrhd6
Jacquelina.Woods , rachel.rodriguez
TITLE: RT-qPCR Detection of Process Controls (Murine noroviurs and crAssphage) from Wastewater using AB 7500 AUTHORS: Jacquelina.Woods , rachel.rodriguez [DESCRIPTION] This method was developed at the FDA’s Center for Food Safety and Applied Nutrition for GenomeTrakr’s pandemic response project, monitoring SARS-CoV...
["[Master Mix Preparation] Prepare Master Mix for all sample and control reactions based on the volumes per 25 µl reaction in this table. Composition of mixes are listed here: Reagent Mixes for RT-qPCR Detection of Process Controls from Wastewater Samples (protocols.io) and should be prepared in advance and stored appr...
87,732
Fluorescent Western Protocol
4
null
https://www.protocols.io/view/fluorescent-western-protocol-czwux7ew
Lynn Doran, Steven J Burgess
TITLE: Fluorescent Western Protocol AUTHORS: Lynn Doran, Steven J Burgess [DESCRIPTION] Analysis of proteins using fluorescent immunoblot with optional total protein stain. Note: - The choice of secondary antibody depends on the choice of primary antibody, whether it is derived from a mouse (monoclonal) or a rabbit...
["Keep membranes in the black Western Blot incubation box for all steps, this is important after adding the secondary antibody because the signal is light-sensitive and will become bleached if exposed to light for a long enough period.", "[Fluorescent Western Protocol] Wet with 1x PBS for 2 min min", "[Fluorescent West...
20,446
Serial Dilution of Nucleofected iPSC Pools
null
dx.doi.org/10.17504/protocols.io.x76frre
null
Celeste Karch, Rita Martinez, Jacob Marsh
TITLE: Serial Dilution of Nucleofected iPSC Pools AUTHORS: Celeste Karch, Rita Martinez, Jacob Marsh [STEPS] ?. Coat 3 wells of a 6 well plate with Matrigel (supplemented with RGD fragment) 1 ml ?. Aspirate media from cells in culture. ?. Wash with - PBS per well. 1 ml 2 ml ?. Add Accutase per well. 1 ml ?. Incubate...
["Coat 3 wells of a 6 well plate with Matrigel (supplemented with RGD fragment)\n1 ml", "Aspirate media from cells in culture.", "Wash with - PBS per well.\n1 ml\n2 ml", "Add Accutase per well.\n1 ml", "Incubate at for to achieve single cells.\nIndividual donor lines exhibit variable sensitivity to accutase-mediated...
98,507
Multiple Myeloma Immune Atlas Consortium: Gene Expression Profiling of the Bone Marrow Microenvironment
0
dx.doi.org/10.17504/protocols.io.q26g718z3gwz/v1
https://www.protocols.io/view/multiple-myeloma-immune-atlas-consortium-gene-expr-dcfj2tkn
Manoj Bhasin
TITLE: Multiple Myeloma Immune Atlas Consortium: Gene Expression Profiling of the Bone Marrow Microenvironment AUTHORS: Manoj Bhasin [DESCRIPTION] A comprehensive gene expression (GEX) profiling protocol that assists in generating the most granular map of single cell landscape from clinical trials supported by Multip...
["[Thawing &amp; Washing bone marrow (BM) derived cells] Preparation", "[GEM Generation and Barcoding] Prepare Master Mix on ice according to the standard 10X protocol and dispense 31.8 µL per sample into a 8 strip tube kept on ice.", "[Sample Index PCR] Choose the appropriate sample index sets to ensure that no sample...
28,004
HiFi Gibson Assembly (Protocol for the NEBuilder® HiFi DNA Assembly Master Mix)
null
dx.doi.org/10.17504/protocols.io.7kchksw
null
Alba Balletbó
TITLE: HiFi Gibson Assembly (Protocol for the NEBuilder® HiFi DNA Assembly Master Mix) AUTHORS: Alba Balletbó [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is the protocol for DNA Assembly using the NEBuilder® HiFi DNA Assembly Master Mix.</div></div> [STEPS] ?. Set up the following reaction...
["Set up the following reaction on ice (to 20μl total volume): ABC12-3 Fragment Assembly4-6 Fragment Assembly2DNA RatioVector:Insert = 1:2Vector:Insert = 1:13Total amount of Fragments0.03-0.2 pmols0.2-0.5 pmols4NEBuilder HiFi DNA Assembly Master Mix10 μL10 μL5Deionized H2O10 - X μL10 - X μL6Total Volume20 μL20 μL\nABC...
82,017
10x Protocols: Visium v2 CytAssist FFPE Library Construction -- University of Minnesota TMCs (CG000495 Rev D)
1
dx.doi.org/10.17504/protocols.io.3byl4jpzzlo5/v1
https://www.protocols.io/view/10x-protocols-visium-v2-cytassist-ffpe-library-con-cub9wsr6
IOx Genomics, Laura J Niedernhofer, David A Bernlohr
TITLE: 10x Protocols: Visium v2 CytAssist FFPE Library Construction -- University of Minnesota TMCs (CG000495 Rev D) AUTHORS: IOx Genomics, Laura J Niedernhofer, David A Bernlohr [DESCRIPTION] Protocols from 10x Genomics for Visium Spatial Gene Expression v2 chemistry on FPPE samples with the CytAssist component. Pro...
["10x protocol CG000 495, Revision D (Library construction with CytAssist)", "Additional Protocols/Resources\nhttps://www.10xgenomics.com/support/spatial-gene-expression-ffpe"]
19,342
High-Density Penetrating Microelectrode Recordings from Anesthetized Feline Dorsal Root Ganglia
1
dx.doi.org/10.17504/protocols.io.w5nfg5e
https://www.protocols.io/view/high-density-penetrating-microelectrode-recordings-w5nfg5e
Zachariah Sperry, Kyounghwan Na, Mihaly Vöröslakos, Saman Parizi, James Jun, Tim M. Bruns, Euisik Yoon, John P. Seymour
TITLE: High-Density Penetrating Microelectrode Recordings from Anesthetized Feline Dorsal Root Ganglia AUTHORS: Zachariah Sperry, Kyounghwan Na, Mihaly Vöröslakos, Saman Parizi, James Jun, Tim M. Bruns, Euisik Yoon, John P. Seymour [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Dorsal root ganglia ...
["[Diamond Shuttle Fabrication]\nPattern tapered trenches onto silicon wafer.", "[Recording Apparatus Assembly]\nBond array interconnects to circuit board for connection to Omnetics adaptor.", "[Insertion and Stabilization Apparatus Assembly]\nMove subject to support stand with overhead rack and ability to lower legs b...
81,504
Preparação para ir ao campo
1
dx.doi.org/10.17504/protocols.io.dm6gpjz31gzp/v2
https://www.protocols.io/view/prepara-o-para-ir-ao-campo-ctt8wnrw
Alicia Rafaelly Vilefort Sales, Daniel Vartanian, Maria Augusta Medeiros Andrade
TITLE: Preparação para ir ao campo AUTHORS: Alicia Rafaelly Vilefort Sales, Daniel Vartanian, Maria Augusta Medeiros Andrade [DESCRIPTION] Este protocolo foi desenhado para o processo de coleta de dados do projeto de pesquisa Associações entre a duração e a qualidade do sono de gestantes no terceiro trimestre com a du...
["[Agenda] Consulte a agenda do projeto e verifique a quantidade de gestantes a serem recrutadas no seu turno.", "[Agenda] Verifique os horários e o trajeto que você terá que fazer para que você chegue com ao menos 30 minutos de antecedência na Casa de Parto.", "[Actígrafos] Separe um actígrafo para cada gestante a ser...
92,647
Use of the waxworm Galleria mellonella larvae as an infection model to study Acinetobacter baumannii
4
dx.doi.org/10.17504/protocols.io.n92ldpr38l5b/v2
https://www.protocols.io/view/use-of-the-waxworm-galleria-mellonella-larvae-as-a-c6qfzdtn
Kah Ern Ten, Nazmul Hasan Muzahid, Sadequr Rahman, tan.hocksiew
TITLE: Use of the waxworm Galleria mellonella larvae as an infection model to study Acinetobacter baumannii AUTHORS: Kah Ern Ten, Nazmul Hasan Muzahid, Sadequr Rahman, tan.hocksiew [DESCRIPTION] Galleria mellonella larvae have been increasingly used in various scientific research, including microbial infection studie...
["[Galleria mellonella rearing and maintenance] Research-grade Galleria mellonella larvae were ordered in bulk from Carolina Biological (US).", "[Galleria mellonella rearing and maintenance] Set up Galleria mellonella housing according to Figure 2.", "[Galleria mellonella rearing and maintenance] Fill 2/3 of the larvae...
102,535
Platelet purification and coating of plates for adhesion assays 
0
dx.doi.org/10.17504/protocols.io.n92ld8wb8v5b/v1
https://www.protocols.io/view/platelet-purification-and-coating-of-plates-for-ad-dgdf3s3n
Samantha King
TITLE: Platelet purification and coating of plates for adhesion assays  AUTHORS: Samantha King [DESCRIPTION] This is the protocol we use to prepare fixed platelets from human blood and coat wells ready for adhesion assays. [GUIDELINES] We typically have greater success when using at least 180 mls of blood. When usi...
["Blood from healthy donors was collected in 1/6th volume of acid citrate dextrose solution (ACD). Mix gently.", "Blood was spun down for 20 min at 200 x g, deaccelerating step without brake.", "Platelet rich plasma was collected (top yellow layer) and mixed (inverting the tube) 1:1 with HEP buffer.", "Platelets were c...
52,645
NCBI submission protocol for SARS-CoV-2 wastewater data: SRA, BioSample, and BioProject
1
dx.doi.org/10.17504/protocols.io.bxndpma6
https://www.protocols.io/view/ncbi-submission-protocol-for-sars-cov-2-wastewater-bxndpma6
Ruth Timme, Maria Balkey
TITLE: NCBI submission protocol for SARS-CoV-2 wastewater data: SRA, BioSample, and BioProject AUTHORS: Ruth Timme, Maria Balkey [DESCRIPTION] PURPOSE: This method was developed at the FDA’s Center for Food Safety and Applied Nutrition for GenomeTrakr’s pandemic response project, monitoring SARS-CoV-2 variants in ...
["[&quot;Ingredients&quot; to have in place before starting your submissions] Set up a new NCBI submission environment for your lab\n1.1: Create an NCBI user account\n1.2: Set up an NCBI submission user group for your lab\n1.4: Bookmark the link to your Submission Portal\n1.5. Identify or establish new BioProjects (det...
54,237
Titan Clear Labs SARS-CoV-2 Strain Characterization Workflow for the Terra Platform
5
null
https://www.protocols.io/view/titan-clear-labs-sars-cov-2-strain-characterizatio-by75pzq6
Frank J Ambrosio, Jill V Hagey, Kevin Libuit, Technical Outreach and Assistance for States Team
TITLE: Titan Clear Labs SARS-CoV-2 Strain Characterization Workflow for the Terra Platform AUTHORS: Frank J Ambrosio, Jill V Hagey, Kevin Libuit, Technical Outreach and Assistance for States Team [DESCRIPTION] The Titan_ClearLabs workflow is a part of the Public Health Viral Genomics Titan series for SARS-CoV-2 geno...
["[Setup Terra and Google Cloud Accounts]", "[Import Titan Clear Labs workflow from Dockstore] Importing the Titan Workflow from Dockstore to the User Workspace\n\nThe Titan Clear Labs workflow is hosted in the Theiagen Dockstore (https://dockstore.org/) repository and has to be imported into the user's Terra Workspace...
79,371
DNA extraction (BOMB)
4
dx.doi.org/10.17504/protocols.io.n2bvj6mdnlk5/v3
https://www.protocols.io/view/dna-extraction-bomb-crrjv54n
Tsu-Chun Hung, Yin-Tse Huang
TITLE: DNA extraction (BOMB) AUTHORS: Tsu-Chun Hung, Yin-Tse Huang [DESCRIPTION] DNA extraction (BOMB) [STEPS] SECTION: Sample Collection 1. Add 200 µL of 1mm beads to 2ml eppendorf tube SECTION: Sample Collection 2. Add200 µL of 0.5mm beads to 2ml eppendorf tube SECTION: Sample Collection 3. Add867 µL Lysis mast...
["[Sample Collection] Add 200 µL of 1mm beads to 2ml eppendorf tube", "[Sample Collection] Add200 µL of 0.5mm beads to 2ml eppendorf tube", "[Sample Collection] Add867 µL Lysis master mix to 2ml eppendorf tube", "[Sample Collection] Collect 10-20 mg of sample to 2ml eppendorf tube", "[Sample crush] Put 2ml eppendorf tu...
19,963
U Mass - Ammonia
null
dx.doi.org/10.17504/protocols.io.xq3fmyn
null
Jason Kim
TITLE: U Mass - Ammonia AUTHORS: Jason Kim [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block">This experiment involves a spectrophotometric measurement using Roche Cobas Clinical Chemistry Analyzer.</div></div> [STEPS] ?....
["Perform daily quality control assessment of instrumentation before analysis.", "Load each sample into a specialized micro-sample cup for the clinical chemistry analyzer.", "Select Ammonia test on display and run the analysis.", "Collect and analyze the data."]
81,675
Individual AAV production and purification
4
dx.doi.org/10.17504/protocols.io.14egn2dqzg5d/v1
https://www.protocols.io/view/individual-aav-production-and-purification-ctzjwp4n
Miguel Chuapoco
TITLE: Individual AAV production and purification AUTHORS: Miguel Chuapoco [DESCRIPTION] We recently developed adeno-associated virus (AAV) capsids to facilitate efficient and noninvasive gene transfer to the central and peripheral nervous systems. However, a detailed protocol for generating and systemically deliverin...
["[Reagent setup: Plasmid DNA] Grow bacterial stocks in LB or Plasmid+ media containing the appropriate selective antibiotic; refer\nto the Addgene catalog for suggested growth conditions. Use a large-scale endotoxin-free plasmid purification kit to isolate plasmids; elute plasmid DNA with the supplied Tris-EDTA (TE) b...
97,520
Tissue H&amp;E Staining | HuBMAP | JHU-TMC
0
dx.doi.org/10.17504/protocols.io.kqdg3242qv25/v1
https://www.protocols.io/view/tissue-h-amp-e-staining-hubmap-jhu-tmc-dbgq2jvw
Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Miklhail James, Sashank Reddy, Denis Wirtz
TITLE: Tissue H&amp;E Staining | HuBMAP | JHU-TMC AUTHORS: Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Miklhail James, Sashank Reddy, Denis Wirtz [DESCRIPTION] The following are guidelines that will have an effective staining window of 2 to 5 minutes. Developed as progressive stains, the intensity of nucl...
["[H&amp;E staining] Final Step: Mount and Coverslip with Covermount", "Set 2 stations with 100% ethanol. Submerge the slide for a minute in each station.", "[H&amp;E staining] Wash the slide in running warm water for a minute", "[H&amp;E staining] Submerge the slide in Optik Hematoxylin for 3 minutes", "[H&amp;E stain...
64,423
Ocuprime Scam Or Legit Updated 2022 Must See before buy
3
dx.doi.org/10.17504/protocols.io.eq2lynqywvx9/v1
https://www.protocols.io/view/ocuprime-scam-or-legit-updated-2022-must-see-befor-ca6fshbn
fluxactive
TITLE: Ocuprime Scam Or Legit Updated 2022 Must See before buy AUTHORS: fluxactive [DESCRIPTION] (LOWEST PRICE GUARANTEED) Click Here to Buy Ocuprime For The Lowest Price Today [STEPS]
[]
64,722
Beneficial Bio Products: Quality control tests 
4
null
https://www.protocols.io/view/beneficial-bio-products-quality-control-tests-cbfssjne
Nadine Mowoh, Stephane Fadanka
TITLE: Beneficial Bio Products: Quality control tests  AUTHORS: Nadine Mowoh, Stephane Fadanka [DESCRIPTION] After production, we typically subject our products to a batch of quality control assays to ascertain their functionality, efficacy and ability to meet their intended purpose. Strict quality control is nec...
["[Functionality tests] Pipetting:\n \n\nDNA polymerase and Master mix enzyme\n\nThaw all reagents on ice in a bowl\nLabel reaction tubes (PCR tubes) according to the number of samples, and including controls in each run (negative and positive controls) as needed.\n\nPolymerase enzyme type:\nIf using a 10x cellular rea...
81,449
Surgical decision-making in the management of knee cartilage injuries in football players: Development of patient-specific recommendations using the RAND/UCLA appropriateness method
1
null
https://www.protocols.io/view/surgical-decision-making-in-the-management-of-knee-ctshwnb6
Giuseppe Filardo, Andreas Serner, Luca Andriolo, Andrew Massey, Emmanuel Papakostas, Peter Verdonk, Elizaveta Kon
TITLE: Surgical decision-making in the management of knee cartilage injuries in football players: Development of patient-specific recommendations using the RAND/UCLA appropriateness method AUTHORS: Giuseppe Filardo, Andreas Serner, Luca Andriolo, Andrew Massey, Emmanuel Papakostas, Peter Verdonk, Elizaveta Kon [DESCRI...
[]