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19,293
Instructions for recreating elPrep 4.0.0 WGS benchmarks
null
dx.doi.org/10.17504/protocols.io.w35fgq6
null
Charlotte Herzeel
TITLE: Instructions for recreating elPrep 4.0.0 WGS benchmarks AUTHORS: Charlotte Herzeel [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Instructions for recreating the elPrep4.0.0 WGS benchmarks used in the following paper:</div><div class = "text-block">Herzeel C, Costanza P, Decap D, Fostier J, ...
["Configuration1.1 Hardware1.2 Software\nThese instructions have been tested with elPrep v.4.0.0. The following assumes that everything is performed from a working directory WORKDIR.\n* 2x18-core Intel Xeon processor E5-2699v3 Haswell @ 2.3GHz* 256 GB RAM* 2x400 GB SSD\n* Ubuntu 14.04.5 LTS* elPrep 4.0.0", "Installatio...
78,871
Kraus et al., 2022 FBXO7 /Park15  
2
dx.doi.org/10.17504/protocols.io.kxygx99pwg8j/v2
https://www.protocols.io/view/kraus-et-al-2022-fbxo7-park15-cq9xvz7n
Felix Kraus
TITLE: Kraus et al., 2022 FBXO7 /Park15   AUTHORS: Felix Kraus [DESCRIPTION] The protein kinase PINK1 and ubiquitin ligase Parkin promote removal of damaged mitochondria via a feed-forward mechanism involving ubiquitin (Ub) phosphorylation, Parkin activation, and ubiquitylation of mitochondrial outer membrane proteins...
[]
92,655
Spinal manipulations for migraine
1
dx.doi.org/10.17504/protocols.io.q26g7pne9gwz/v1
https://www.protocols.io/view/spinal-manipulations-for-migraine-c6qpzdvn
Paul Posadzki, Edzard Ernst
TITLE: Spinal manipulations for migraine AUTHORS: Paul Posadzki, Edzard Ernst [DESCRIPTION] Several new randomized trials evaluating the effectiveness and safety of spinal manipulative therapies (SMT) have been published since our previous systematic review (SR). Therefore, we aim at updating the evidence base for the...
["Spinal manipulations for migraine: a protocol for an updated systematic review and meta-analysis of randomized clinical trials", "[Protocol]"]
103,899
Effect of OT-Bridge System Versus Multiunit Abutment on the Passive Fit and Stresses Induced by CAD/CAM Fabricated All-on-Four Screw-Retained Prostheses (In Vitro Study)
0
dx.doi.org/10.17504/protocols.io.14egn6e4zl5d/v1
https://www.protocols.io/view/effect-of-ot-bridge-system-versus-multiunit-abutme-dhp335qn
Abdelrahman Hazem, Shaimaa Lotfy Mohamed, Sara Ibrahim Soliman Mohamed
TITLE: Effect of OT-Bridge System Versus Multiunit Abutment on the Passive Fit and Stresses Induced by CAD/CAM Fabricated All-on-Four Screw-Retained Prostheses (In Vitro Study) AUTHORS: Abdelrahman Hazem, Shaimaa Lotfy Mohamed, Sara Ibrahim Soliman Mohamed [DESCRIPTION] The hazardous effect of stresses induced by lack...
["[Abstract] The hazardous effect of stresses\ninduced by lack of passivity in screw-retained prostheses necessitates the\nselection of a suitable intermediate component between the implant fixture and\nthe prosthetic framework. Thus, this study aims to evaluate passivity and\nstresses induced by CAD/CAM screw-retained...
22,493
The Illumina libraries preparation for the Scapharca broughtonii
null
dx.doi.org/10.17504/protocols.io.z75f9q6
null
Chang-Ming Bai
TITLE: The Illumina libraries preparation for the Scapharca broughtonii AUTHORS: Chang-Ming Bai [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol is used to detail the process of Illumina libraries preparation for the</span><span style = "font-style:italic;"> Scapharca broughtonii<...
["The extracted DNA was sheared into fragments about 350 bp in size using a Bioruptor Pico Sonication System (Diagenode, Seraing, Belgium), and verified by agarose gel electrophoresis.\nThe sizes of the main fragments should between 200 bp and 500 bp.", "Fragments with size > 300 bp were purified using VAHTSTM DNA Clea...
94,641
Scoping Review: Malaria risk stratification and incidence mapping
1
dx.doi.org/10.17504/protocols.io.3byl4qn9zvo5/v1
https://www.protocols.io/view/scoping-review-malaria-risk-stratification-and-inc-c8nrzvd6
Jessica Craig, Donal Bisanzio, Richard Reithinger
TITLE: Scoping Review: Malaria risk stratification and incidence mapping AUTHORS: Jessica Craig, Donal Bisanzio, Richard Reithinger [DESCRIPTION] Context: As countries continue to make progress against malaria at national level, transmission of malaria and, hence, risk becomes increasingly heterogeneous. To maximize a...
["[Rationale] Malaria is an acute febrile illness caused by a parasitic\ninfection transmitted by Anopheles mosquitoes. Human malaria is caused by five\ndifferent Plasmodium parasites, with P. falciparum being the predominant\nspecies in sub-Saharan Africa (SSA) [1]. Malaria burden has a marked spatial\nheterogeneity w...
84,080
DNA/RNA extraction from fresh-frozen tissue, AllPrep DNA/RNA/miRNA Universal Kit
4
null
https://www.protocols.click/view/dna-rna-extraction-from-fresh-frozen-tissue-allpre-cwcqxavw
Annika Fendler
TITLE: DNA/RNA extraction from fresh-frozen tissue, AllPrep DNA/RNA/miRNA Universal Kit AUTHORS: Annika Fendler [DESCRIPTION] Protocol for combined RNA and DNA extraction from fresh-frozen tissue using the AllPrep DNA/RNA/miRNA Universal Kit. [BEFORE_START] Preparations: FRN buffer: Add 42 ml Isoprop to new bottle RP...
["[Tissue preparation] This protocol is for \nOptional: Weigh tissue", "[Tissue preparation] Transfer tissue in 350 µL 600 µL RLT + ß-ME in 2 ml DNA LoBind tube.\n\nMake sure that the tissue does not defrost.", "[Tissue preparation] Add a 5mm bead to each tube and lyse tissue in TissueLyser for 2 min @ 20 Hz", "[T...
27,385
FTÅ200 Interfacial Tension Measurement
1
null
https://www.protocols.io/view/ft-200-interfacial-tension-measurement-6yzhfx6
Kenneth Schackart
TITLE: FTÅ200 Interfacial Tension Measurement AUTHORS: Kenneth Schackart [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">How to use the First Ten Ångstrom (FTÅ) 200 for measuring interfacial tension of aqueous solution. Some aspects of this protocol are specific to use in Yoon Biosensors Lab at The ...
["[Mount Syringe]\nTwist needle onto glass syringe until tight, but not excessively tight.\nUse caution when grabbing needle. Avoid touching the tips of needles to prevent contamination.", "[Mount Syringe]\nMount syringe by sliding through the hole on the syringe mount, bringing the two halves of the white syringe clam...
81,800
Preparação para ir ao campo
1
dx.doi.org/10.17504/protocols.io.dm6gpjz31gzp/v3
https://www.protocols.io/view/prepara-o-para-ir-ao-campo-ct5gwq3w
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 Sono e Gravidez. Consulte a aba Guidelines para visualizar o fluxograma do processo como um t...
["[Agenda] Consulte a agenda do projeto e verifique a quantidade de gestantes a serem recrutadas em 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 do horário de sua primeira consulta na Casa de Parto.", "[Actígrafos] Separe u...
43,820
Single-step generation of genetic reporter lines using CRISPR/Cas9 and transient antibiotic selection
4
null
https://www.protocols.io/view/single-step-generation-of-genetic-reporter-lines-u-bn2kmgcw
Carles Calatayud, William Serkin, Patrik Verstreken
TITLE: Single-step generation of genetic reporter lines using CRISPR/Cas9 and transient antibiotic selection AUTHORS: Carles Calatayud, William Serkin, Patrik Verstreken [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>The following protocol describes the generation of genetic reporters in hum...
["[Preparing Cas9 mRNA and the gene-specific guide RNA through in vitro transcription (IVTR)]\nAmplify Cas9-T2A-mClover3 IVTR template from pIVTR(T7A)-Cas9-T2A-mClover3 using the following primers AB1Pre_T7_FwdACATTTCCCCGAAAAGTGCC2HBB_3UTR_150T_RvTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT...
22,334
Transfection of Crypthecodinium cohnii using labelled DNA
null
dx.doi.org/10.17504/protocols.io.z26f8he
null
José A. Fernández Robledo
TITLE: Transfection of Crypthecodinium cohnii using labelled DNA AUTHORS: José A. Fernández Robledo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">José A. Fernández Robledo </div><div class = "text-block">Nastasia J. Freyria</div><div class = "text-block"> Bigelow Laboratory for Ocean Sciences</div...
["[Reagents]\nLiquid mediumL1 + 10 gr/L glucose NPN (https://ncma.bigelow.org/ccmp316#.XLOpDZNKi2w)L1 medium with nitrogen limited (without yeast extract and glucose) to obtain cyst from C. cohnii swimming cellsPlatesPrepare Petri dishes containing L1 medium bacteriological agar (1.5%)Prepare PEG 8000 solution (0.4%) ...
71,099
SHInE - Simultaneous HCR, Immunohistochemistry, Nuclear staining and EdU
4
dx.doi.org/10.17504/protocols.io.5qpvobnyzl4o/v1
https://www.protocols.io/view/shine-simultaneous-hcr-immunohistochemistry-nuclea-chn3t5gn
Aida Coric, Alexander W. Stockinger, Petra Schaffer, Dunja Rokvic, Kristin Tessmar-Raible, Florian Raible
TITLE: SHInE - Simultaneous HCR, Immunohistochemistry, Nuclear staining and EdU AUTHORS: Aida Coric, Alexander W. Stockinger, Petra Schaffer, Dunja Rokvic, Kristin Tessmar-Raible, Florian Raible [DESCRIPTION] This protocol allows the multiplexed use of four different molecular labelling techniques in whole-mount Platy...
["[Optional: EdU incubation] Optional: Chase out EdU with 3 millimolar (mM) thymidine for 60 min ; then replace thymidine solution with fresh ASW and incubate for desired chase length", "[Optional: EdU incubation] Incubate worms with 10 micromolar (µM) EdU in artificial sea water (ASW) (EdU aliquots in both DMSO and H...
null
null
null
dx.doi.org/10.17504/protocols.io.pnpdmdn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><u>Goal:</u></p> <p>This document aims to standardize the protocol for assaying the association of bacteria or fluorescent latex beads in eukaryotic cells by flow cytometry.</p> <p> </p> <p><u>General considerations:</u></p> <p>- There are different models of flow cytometers,...
[]
97,130
Data-Independent Acquisition (DIA) Data Processing using Spectronaut/directDIA (Biognosys): Whole Proteome Analysis of Human Whole Lung Tissue
0
dx.doi.org/10.17504/protocols.io.261ge5p8og47/v1
https://www.protocols.io/view/data-independent-acquisition-dia-data-processing-u-da4i2gue
Joanna Bons, J P Rose, M A Watson, B Schilling
TITLE: Data-Independent Acquisition (DIA) Data Processing using Spectronaut/directDIA (Biognosys): Whole Proteome Analysis of Human Whole Lung Tissue AUTHORS: Joanna Bons, J P Rose, M A Watson, B Schilling [DESCRIPTION] Proteolytic peptide measurement of Human whole lung tissue using Liquid Chromatography-Tandem Mass ...
["Search the DIA data with Spectronaut using the spectral library-free directDIA\nalgorithm against a Human UniProt-SwissProt proteome database (https://www.uniprot.org/proteomes), applying the settings described in Table 1."]
30,640
Visual attentional tasks
null
dx.doi.org/10.17504/protocols.io.96qh9dw
null
Chiara Valeria Marinelli, Pierluigi Zoccolotti, Cristina Romani
TITLE: Visual attentional tasks AUTHORS: Chiara Valeria Marinelli, Pierluigi Zoccolotti, Cristina Romani [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The tasks examined a) visuo-spatial memory; b) visuo-attentional capacity; and c) serial visual-attention.</div></div> [STEPS]
[]
21,129
Preparation of human Red Blood Cells for Scanning Electron Microscopy imaging
null
dx.doi.org/10.17504/protocols.io.yvhfw36
null
Marie Anne Balanant
TITLE: Preparation of human Red Blood Cells for Scanning Electron Microscopy imaging AUTHORS: Marie Anne Balanant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">SEM imaging can be used to quantify RBC shape. RBCs are sampled from fresh blood samples or from packed RBC units during storage. RBCs are...
["[Coverslip preparation]\nPrepare glass coverslips by incubating them in poly-D-lysine solution in PBS for at RT\n10 Volume Percent", "[Coverslip preparation]\nRinse twice with milliQ water", "[Coverslip preparation]\nDry and keep at until use", "[RBC fixation with glutaraldehyde]\nWash of RBC in PBS ()\n100 µl\n...
null
null
null
dx.doi.org/10.17504/protocols.io.shheb36
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol refers to a study having the aim to identify and test at field artificial structures suitable as collectors for common cuttlefish Sepia officinalis eggs in wild condition.</p> <p>During the same study, three different protocols were tested and compared. This one...
[]
50,155
Sample collection and DNA purification from virus-enriched tangential flow filtration retentates
1
null
https://www.protocols.io/view/sample-collection-and-dna-purification-from-virus-bu8jnzun
Paul Den Uyl, Ed Delong
TITLE: Sample collection and DNA purification from virus-enriched tangential flow filtration retentates AUTHORS: Paul Den Uyl, Ed Delong [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">To collect and concentrate viral particles, seawater samples were collected and prefiltered to remove bacterioplan...
["[Pre-Cleaning]\nRecirculate 0.1N Sodium Hydroxide at feed speed for .\nCentrifuge: 330 33", "[Pre-Cleaning]\nFlush with MilliQ.\n30 L", "[Pre-Cleaning]\nDrain the system (see sample recovery).", "[Concentrating]\nFill retentate reservoir.", "[Concentrating]\nAttach retentate reservoir.", "[Concentrating]\nConnect f...
21,008
Environmental DNA (eDNA) extraction with modified Qiagen DNeasy Blood & Tissue kit
null
dx.doi.org/10.17504/protocols.io.yrqfv5w
null
Collin Closek, Hilary Starks, Elizabeth Andruszkiewicz, Alexandria Boehm
TITLE: Environmental DNA (eDNA) extraction with modified Qiagen DNeasy Blood & Tissue kit AUTHORS: Collin Closek, Hilary Starks, Elizabeth Andruszkiewicz, Alexandria Boehm [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Nucleic acids extraction from the filters using the Qiagen DNeasy Blood and Tiss...
["[Preparation]\nNucleic acids were extracted from the filters using the Qiagen DNeasy Blood and Tissue Kit with some modifications to the manufacturer’s protocol.", "[Preparation]\nPrior to extraction, 0.5 mm and 0.1 mm glass beads (BioSpec Products) transferred to glass beaker, covered with tin foil, and ashed at 500...
106,332
Pollen germination on wheat stigmas
0
dx.doi.org/10.17504/protocols.io.kxygxy42ol8j/v1
https://www.protocols.io/view/pollen-germination-on-wheat-stigmas-dj344qqw
Marina Millán Blánquez
TITLE: Pollen germination on wheat stigmas AUTHORS: Marina Millán Blánquez [DESCRIPTION] Aniline blue staining of pollen tube growth on the wheat stigma and through the style 4 h and 30 min after pollination. [STEPS] SECTION: Hand pollination 1. Please refer to wheat-training.com for detailed explanations on how to ...
["[Hand pollination] Please refer to wheat-training.com for detailed explanations on how to perform hand pollinations. Link to pdf: https://www.wheat-training.com/wp-content/uploads/Wheat_growth/pdfs/How-to-cross-wheat-pdf.pdf", "[Carpel dissection and fixation] Using a pair of tweezers, dissect carpels 4.5 h after pol...
41,618
Harmony Transfer Pipette Protocol
4
dx.doi.org/10.17504/protocols.io.bkvskw6e
https://www.protocols.io/view/harmony-transfer-pipette-protocol-bkvskw6e
Jack H Kotnik, Nuttada Panpradist, Ian Hull, Qin Wang, Mike Roller, Amy Oreskovic, Enos Kline, Daniel Leon, Barry Lutz, Robert Atkinson
TITLE: Harmony Transfer Pipette Protocol AUTHORS: Jack H Kotnik, Nuttada Panpradist, Ian Hull, Qin Wang, Mike Roller, Amy Oreskovic, Enos Kline, Daniel Leon, Barry Lutz, Robert Atkinson [STEPS] ?. [Intended Use] The Harmony assay is used with the Harmony instrument for the detection of SARS-CoV-2 viral RNA in lower na...
["[Intended Use]\nThe Harmony assay is used with the Harmony instrument for the detection of SARS-CoV-2 viral RNA in lower nares (nasal swab) specimens collected from individuals suspected of SARS-CoV-2 infection. The assay is intended for the qualitative detection of SARS-CoV-2 nucleic acid from direct specimens.The a...
null
null
null
dx.doi.org/10.17504/protocols.io.mahc2b6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Efficiency of plaquing (EOP) assay for <em>C. jejuni </em>phages</p> <p>Jessica Sacher (with help from Stephanie Guerra</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
32,653
Enzymatic disaggregation of human myometrium for 10X Single Cell RNA-seq
null
dx.doi.org/10.17504/protocols.io.bb5miq46
null
Aymara Mas, Alba Machado-Lopez, Patricia Escorcia, Carlos Simón
TITLE: Enzymatic disaggregation of human myometrium for 10X Single Cell RNA-seq AUTHORS: Aymara Mas, Alba Machado-Lopez, Patricia Escorcia, Carlos Simón [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol describes the procedure for dissociating myometrial samples, which is based in ...
["[Tissue collection and transportation]\nUterine tissues should be transported in preservation solution (HypoThermosol® FRS) at from the surgery room to the lab.\n4 °C", "[Dissociation]\nRinse the sample with wash buffer, removing blood or mucus.", "[Remove red blood cells]\n[Optional] If after centrifugation there i...
88,358
18S V9 PCR
4
null
https://www.protocols.io/view/18s-v9-pcr-c2ieycbe
Kathleen Pitz
TITLE: 18S V9 PCR AUTHORS: Kathleen Pitz [DESCRIPTION] This protocol is aimed at amplifying the 18S rRNA hypervariable region 9 (18S V9) in eukaryotes with a focus on microbial eukaryotes. Amplicons generated using this protocol can then be sequenced using the Illumina platform. The primers (1391F, EukBr) utilized in ...
["[Minimum Information about an Omics Protocol (MIOP)] MIOP Term\nValue\n \nmethodology category\n\n \nproject\n\n \npurpose\n\n \nanalyses\n\n \ngeographic location\n\n \nbroad-scale environmental context\n\n \nlocal environmental context\n\n \nenvironmental medium\n\n \ntarget\n18S gene\n \ncreator\n\n \nmaterials ...
64,396
True Ketogenics Keto Bundle: Weight Loss Pills [Pros &amp; Cons]| Ingredients, Price and Warnings!!
3
dx.doi.org/10.17504/protocols.io.bp2l61y61vqe/v1
https://www.protocols.io/view/true-ketogenics-keto-bundle-weight-loss-pills-pros-ca5ksg4w
shinelsom
TITLE: True Ketogenics Keto Bundle: Weight Loss Pills [Pros &amp; Cons]| Ingredients, Price and Warnings!! AUTHORS: shinelsom [DESCRIPTION] "True Ketogenics Keto Bundle [STEPS]
[]
36,798
Human Neurosurgical Tissue Processing
null
dx.doi.org/10.17504/protocols.io.bf66jrhe
https://www.protocols.io/view/human-neurosurgical-tissue-processing-bf66jrhe
Allen Institute for Brain Science
TITLE: Human Neurosurgical Tissue Processing AUTHORS: Allen Institute for Brain Science [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes the procedure for receiving human surgical tissue, followed by trimming, mounting, embedding, and slicing under sterile conditions in order ...
[]
93,070
Culture of human epithelial cells (skin, cornea, thymus) on 3T3J2 feeder layer cells
4
dx.doi.org/10.17504/protocols.io.14egn31ezl5d/v2
https://www.protocols.io/view/culture-of-human-epithelial-cells-skin-cornea-thym-c65nzg5e
Pierluigi Giuseppe Manti
TITLE: Culture of human epithelial cells (skin, cornea, thymus) on 3T3J2 feeder layer cells AUTHORS: Pierluigi Giuseppe Manti [DESCRIPTION] This document is of vital importance for scientists attempting the culture of human keratinocytes derived from skin, cornea, thymus, esophagus etc. Please strictly adhere to the g...
["[3T3J2 Growth Medium] Open DMEM bottle (ThermoFisher Scientific).", "[3T3J2 Growth Medium] Add 8 % (v/v) of bovine serum (ThermoFisher Scientific) (DO NOT use fetal calf serum!).", "Set up one flask at low density to maintain the line (5*10^5 cells at early passages, 10^5 cells at late passages). This flask should r...
106,319
Methods for xenograft tissue processing and capture for spatial transcriptomics using 10x-Genomics Visium platform
0
dx.doi.org/10.17504/protocols.io.4r3l2q1yxl1y/v1
https://www.protocols.io/view/methods-for-xenograft-tissue-processing-and-captur-dj3p4qmn
Sandy S. Pineda, Hongyun Li, YuHong Fu, Glenda Halliday
TITLE: Methods for xenograft tissue processing and capture for spatial transcriptomics using 10x-Genomics Visium platform AUTHORS: Sandy S. Pineda, Hongyun Li, YuHong Fu, Glenda Halliday [DESCRIPTION] This protocol details the process of preparing grafted mouse brain samples for spatial transcriptomics using 10X Genom...
["[Tissue preparation] Selected 4% PFF-fixed & paraffin-embedded (FFPE) blocks of the xenograft mouse brains at the striatum level were cut in a rotary microtome (HistoCore MULTICUT R Rotary, Leica Biosystems) at 6µm thickness.\nFour tissue sections were cut and placed on each poly-prep slides (Millipore-Sigma, P0425)....
84,692
snm3C-seq3
1
dx.doi.org/10.17504/protocols.io.kqdg3x6ezg25/v1
https://www.protocols.io/view/snm3c-seq3-cwxuxfnw
Anna Bartlett, Rosa Castanon, Hanqing Liu, Yi Zhang, Chongyuan Luo, Joseph R. Ecker
TITLE: snm3C-seq3 AUTHORS: Anna Bartlett, Rosa Castanon, Hanqing Liu, Yi Zhang, Chongyuan Luo, Joseph R. Ecker [DESCRIPTION] We developed a single-nucleus multiomic sequencing technology (snm3C-seq3) to assay both the methylome and chromatin contacts from within the same cell.  We further adapted this protocol to allo...
["[Prepare Single Nuclei] Prepare stock solutions, stored at 4°C\n\n 1. NIM: Sucrose (250 mM), KCl (25 mM), MgCl2 (5 mM), Tris-Cl pH 8.0 (10 mM)\n 2. Dilutent: Tris-Cl pH 8.0 (120 mM), KCl (150 mM), MgCl2 (30 mM)", "[Prepare Single Nuclei] Prepare the following solutions fresh before each experiment\n\n1....
58,578
Well-tempered Metadynamics protocol
5
null
https://www.protocols.io/view/well-tempered-metadynamics-protocol-b5fsq3ne
Vidya Niranjan, Akshay Uttarkar
TITLE: Well-tempered Metadynamics protocol AUTHORS: Vidya Niranjan, Akshay Uttarkar [DESCRIPTION] Metadynamics is a technique in which the potential for one or more chosen variables ("collective variables") is modified by periodically adding a repulsive potential of Gaussian shape at the location given by particula...
["Simulation system setup. \nIn the solvation tab\nThe complex is solvated in an orthorombhic box with the dimensions 10Å additional to the total size of the receptor complex.\nClick on minimize volume to obtain the optimal box dimensions.\n\n \n\nIn the Ions tab\nPlace ions based on the requirement of neutral, acidic...
38,508
Addgene Covid-19, Back To Work Training
3
dx.doi.org/10.17504/protocols.io.bhukj6uw
https://www.protocols.io/view/addgene-covid-19-back-to-work-training-bhukj6uw
Joanne Kamens, Addgene The Nonprofit Plasmid Repository
TITLE: Addgene Covid-19, Back To Work Training AUTHORS: Joanne Kamens, Addgene The Nonprofit Plasmid Repository [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Disclaimer</span><span>: The SOP presented here has been designed by the Addgene nonprofit plasmid reposi...
[]
73,145
SPOT DNA Extraction from 142mm A/E 1µm Filters
1
dx.doi.org/10.17504/protocols.io.ewov1oqeklr2/v1
https://www.protocols.io/view/spot-dna-extraction-from-142mm-a-e-1-m-filters-cjnzumf6
Yi-Chun Yeh, Erin B Fichot, Alma E Parada
TITLE: SPOT DNA Extraction from 142mm A/E 1µm Filters AUTHORS: Yi-Chun Yeh, Erin B Fichot, Alma E Parada [DESCRIPTION] Reference: Lie et al, 2013. Small-scale temporal and spatial variations in protistan community composition at the San Pedro Ocean Time-series station off the coast of southern California. Aquatic Mic...
["[Day1] Prepare a 70°C water bath.", "[Day1] Prepare a razor blade, forceps, and a metal slab.", "[Day1] Cut the 142mm A/E filter into pieces on the metal slab and place it into a 50 ml falcon tube.", "[Day1] Add 6.2 ml of 2X lysis buffer and 500 µl of 0.5 mm zirconia/silica beads.", "[Day1] Bead-beat (vortex) for 1 m...
19,481
Parhyale hawaiensis embryo single cell dissociation
null
dx.doi.org/10.17504/protocols.io.w9zfh76
null
Leo Blondel
TITLE: Parhyale hawaiensis embryo single cell dissociation AUTHORS: Leo Blondel [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Coat all gear with BSA] Prepare 1% BSA solution in ddH2OCoat 1 or more wells from 12 well plateCoat pasteur pipettes for embryo transfersIf using syringes for loading coat tubing...
["[Coat all gear with BSA]\nPrepare 1% BSA solution in ddH2OCoat 1 or more wells from 12 well plateCoat pasteur pipettes for embryo transfersIf using syringes for loading coat tubing and syringes", "[Collect embryos]\nThis protocol was tested on embryos at 12hpf to 72hpf.From the collected females, harvest embryos in t...
95,212
Intact and clean nuclei isolation from wheat meristems
4
dx.doi.org/10.17504/protocols.io.yxmvm3renl3p/v1
https://www.protocols.io/view/intact-and-clean-nuclei-isolation-from-wheat-meris-c88kzzuw
Isabel Faci, Maximillian RW Jones, Cristobal Uauy
TITLE: Intact and clean nuclei isolation from wheat meristems AUTHORS: Isabel Faci, Maximillian RW Jones, Cristobal Uauy [DESCRIPTION] This protocol describes how to perform nuclei isolation from wheat meristem tissues (spikes and carpels/ovaries), specifically from flash-frozen tissues. With this protocol, we obtain ...
["[Tissue collection] Next, for each sample, label 2 mL Eppendorf tubes for collection and place two nitrogen-cooled metallic steel beads (previously cleaned with 0.5 M HCl and 70% ethanol, diameter 3 mm) in each tube. For each sample, we will pool a number of meristematic spikes/ovaries/spikelets. Depending on the sta...
87,061
Standard Operating Procedure (SOP) for Combined ICA and Systemic Administration of MPTP: The Overlesioned (Bilateral Asymmetric) Non-human Primate Model
4
dx.doi.org/10.17504/protocols.io.dm6gp3zw5vzp/v1
https://www.protocols.io/view/standard-operating-procedure-sop-for-combined-ica-cy9vxz66
Robert S Turner
TITLE: Standard Operating Procedure (SOP) for Combined ICA and Systemic Administration of MPTP: The Overlesioned (Bilateral Asymmetric) Non-human Primate Model AUTHORS: Robert S Turner [DESCRIPTION] This standard operating procedure (SOP) describes safe methods for the use of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridi...
["[Receipt and storage of MPTP] Place the package with MPTP into a chemical fume hood.", "[Receipt and storage of MPTP] Don appropriate personal protective equipment (Tyvek suit, half-mask respirator, bonnet, face shield, 2 pairs of nitrile gloves) and inspect the package to ensure no damage.", "[Receipt and storage of...
null
null
null
dx.doi.org/10.17504/protocols.io.dak2cv
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <p><strong>Primers:</strong></p> <p>rpoC1_103:  CAC TGC GGC AAA TAC AAA CG</p> <p>rpoC1_360:  TTT CAG GTC CTT GTG GTC AC<br /><br /></p> <strong><strong>Reaction Mix:<br /></strong></strong> <table style="width: 400px; height: 150px;"> <tbody> <tr> <td>Material</td> <td>Amt.</td>...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.rh6d39e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol for preparing NGFR-mNG2 tagged WM989 A6-G3 melanoma cells for sorting. </p> [BEFORE_START] <p>Warm trypsin, media and DPBS. </p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
67,277
BeLiv [Official Website] 2022
1
dx.doi.org/10.17504/protocols.io.8epv598qjg1b/v1
https://www.protocols.io/view/beliv-official-website-2022-cdxms7k6
sonoviveget
TITLE: BeLiv [Official Website] 2022 AUTHORS: sonoviveget [DESCRIPTION] Many individuals with diabetes battle with fluctuating glucose levels. Diabetes medications and insulin shots can help. Notwithstanding, certain individuals with diabetes search for regular choices to help glucose. As indicated by the US-based p...
[]
79,007
Standard M9 minimal medium
4
null
https://www.protocols.io/view/standard-m9-minimal-medium-crd7v29n
Andreas Sagen
TITLE: Standard M9 minimal medium AUTHORS: Andreas Sagen [DESCRIPTION] M9 minimal media is a highly-referenced microbial growth medium used for the cultivation of E. coli. This buffered minimal microbial medium contains only salts and nitrogen, so it is traditionally supplemented with glucose, amino acids and vitamins...
["[1 000 mL 10x M9 salt solution] In a bottle, fill with 800 mL deionized water", "[1 000 mL 10x M9 salt solution] Measure and add:\n 60 g Sodium phosphate (dibasic)\n 30 g Monopotassium phosphate\n 5 g Sodium chloride\n\nMaterials:", "[1 000 mL 10x M9 salt solution] Adjust pH to pH 7.4 using hydrogen chloride...
47,416
Automated QIAcube DNA Extraction
4
dx.doi.org/10.17504/protocols.io.5qpvoypkdg4o/v1
https://www.protocols.io/view/automated-qiacube-dna-extraction-bsiyncfw
Clemens Scherzer
TITLE: Automated QIAcube DNA Extraction AUTHORS: Clemens Scherzer [DESCRIPTION] This protocol explains the Standard Operating Protocol for automated extraction of DNA using QIAcube. [BEFORE_START] Based on 12 Samples. DNA Q/C GOALS 1. Cary Concentration Assay a. 260/280 = 1.8-2.0 b. Manual Purag...
["[Part A: First Elution] Heat water bath to 37 °C before starting.", "[Part A: First Elution] Thaw Buffy Coat samples by gently agitating sample in water bath.", "[Part A: First Elution] Turn on QIAcube. Open door and remove reagent tray from QIAcube. Unscrew all caps and refill reagents where necessary. Do not exceed...
77,397
Virome harvesting from cheese microbiomes
4
dx.doi.org/10.17504/protocols.io.j8nlkw29xl5r/v1
https://www.protocols.io/view/virome-harvesting-from-cheese-microbiomes-cptvvnn6
Adair Borges
TITLE: Virome harvesting from cheese microbiomes AUTHORS: Adair Borges [DESCRIPTION] This protocol details how we harvest the viral component, or virome, from cheese rind microbiomes. These viromes can be banked at 4 °C for short-term storage or -80 °C for long-term storage. Alternatively, one can extract DNA or RNA ...
["[Virome extraction from cheese microbial community] Once the cheese sample is fully suspended, add 1 mL of chloroform to the sample. Close the conical tube, and seal the top with Parafilm. Shake the sample on a platform shaker for 1–2 h at room temperature.", "[Virome extraction from cheese microbial community] Move ...
78,314
Getting started with Micro-Meta App Tutorial
5
dx.doi.org/10.17504/protocols.io.36wgq7ddxvk5/v7
https://www.protocols.io/view/getting-started-with-micro-meta-app-tutorial-cqqivvue
Alessandro Rigano, Mathias Hammer, Joel Ryan, Claire M. Brown, David Grunwald, Caterina Strambio De Castillia
TITLE: Getting started with Micro-Meta App Tutorial AUTHORS: Alessandro Rigano, Mathias Hammer, Joel Ryan, Claire M. Brown, David Grunwald, Caterina Strambio De Castillia [DESCRIPTION] For quality, interpretation, reproducibility, and sharing value, microscopy images should be accompanied by detailed descriptions of...
["[Before the tutorial] Video introduction", "[Tutorial - 1 - Download and Install Micro-Meta App] Download Micro-Meta App\n\nFollow the instructions in this step and in Video 3 of the tutorial series to download and install the stand-alone version of the Micro-Meta App.", "[Tutorial - 1 - Download and Install Micro-Me...
37,900
Dephosphorylation of 5´-ends of DNA using rSAP (M0371)
1
dx.doi.org/10.17504/protocols.io.bg9kjz4w
https://www.protocols.io/view/dephosphorylation-of-5-ends-of-dna-using-rsap-m037-bg9kjz4w
New England Biolabs
TITLE: Dephosphorylation of 5´-ends of DNA using rSAP (M0371) AUTHORS: New England Biolabs [DESCRIPTION] Protocol for Dephosphorylation of 5´-ends of DNA using rSAP (M0371). [GUIDELINES] Dephosphorylation of 5´- ends of DNA in Restriction Enzyme Reaction The phosphate can be added directly into the digestion react...
["Prepare a 20 µL reaction as follows: \nDNA1 pmol of DNA ends*CutSmart® Buffer (10X)2 µlrSAP1 unitH2O, purifiedto 20 µl**", "Incubate at 37 °C for 30 min.", "Stop reaction by heat-inactivation at 65 °C for 5 min."]
42,940
Creating sci-Space Grids for Spatial Barcoding
1
null
https://www.protocols.io/view/creating-sci-space-grids-for-spatial-barcoding-bm64k9gw
Sanjay Srivatsan, Mary Regier
TITLE: Creating sci-Space Grids for Spatial Barcoding AUTHORS: Sanjay Srivatsan, Mary Regier [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Sci-Space is a technology for transferring spatially arrayed oligonucleotides onto tissue sections prior to single cell sequencing. The spatially arrayed oli...
["[Casting dried agarose slides]\nTo prepare nuclease-free agarose, 3% w/v (SeaPlaque, Lonza, Bend, OR) was added to deionized water containing 0.1% v/v , incubated 2 hr at room temperature, and autoclaved at 15 psi on liquid cycle for at least 15 min to inactivate the remaining diethyl pyrocarbonate.", "[Casting drie...
18,761
Detection of single nucleotide polymorphisms with padlock probes
null
dx.doi.org/10.17504/protocols.io.wjhfcj6
null
Tomasz Krzywkowski, Mats Nilsson
TITLE: Detection of single nucleotide polymorphisms with padlock probes AUTHORS: Tomasz Krzywkowski, Mats Nilsson [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for in situ detection of mRNAs and single nucleotide polymorphisms using padlock probes and rolling circle amplification....
["[Specimen Pretreatment: Adherent Cells]\nCulture cells in a flask until 80-90% confluence.", "[Specimen Pretreatment: Adherent Cells]\nWash cells twice with PBS, and dislodge cells by adding 1 mL/25 cm2 of 0.25% Trypsin-EDTA for 1-2 min.\n[Trypsin]", "[Specimen Pretreatment: Adherent Cells]\nResuspend the cells in ap...
96,918
Generation of knockout iPSCs using CRISPR-Cas9 genome editing
0
dx.doi.org/10.17504/protocols.io.36wgqnr33gk5/v1
https://www.protocols.io/view/generation-of-knockout-ipscs-using-crispr-cas9-gen-davw2e7e
Nisha Mohd Rafiq, Pietro De Camilli
TITLE: Generation of knockout iPSCs using CRISPR-Cas9 genome editing AUTHORS: Nisha Mohd Rafiq, Pietro De Camilli [DESCRIPTION] This protocol describes the genetic modification of induced pluripotent cells (iPSCs) using CRISPR-Cas9, including synthesis of gRNA plasmids, transfection, selection of clones, and sequencin...
["[Molecular cloning] Design gRNAs using CHOPCHOP (https://chopchop.cbu.uib.no/). Specific gRNAs used in this study can be found in the method section of our manuscript.", "[Molecular cloning] Order gRNAs as complementary single stranded oligonucleotides (Integrated DNA Technologies). The indicated overhanging nucleoti...
null
null
null
dx.doi.org/10.17504/protocols.io.kjecuje
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
36,013
Flow cytometry trait measurements (size, granularity and chlorophyll-a) of diatoms
1
dx.doi.org/10.17504/protocols.io.dm6gpr2x5vzp/v1
https://www.protocols.io/view/flow-cytometry-trait-measurements-size-granularity-bfemjjc6
Phoebe Argyle, Jana Hinners, Nathan Walworth, Sinead Collins, Naomi M. Levine, Martina Doblin
TITLE: Flow cytometry trait measurements (size, granularity and chlorophyll-a) of diatoms AUTHORS: Phoebe Argyle, Jana Hinners, Nathan Walworth, Sinead Collins, Naomi M. Levine, Martina Doblin [DESCRIPTION] A method and guide for measuring size, relative granularity, and chlorophyll-a content in centric diatom cells. ...
["[Sample preparation] Aliquot 200 µL of live, uniform culture and transfer to a round-bottom tissue culture plate. Agitate culture gently to suspend cells to create a uniform culture.", "[Sample preparation] Fix the cells with 20 µL (10 % volume) paraformaldehyde 8% solution, resulting in a final concentration of 0....
null
null
null
dx.doi.org/10.17504/protocols.io.hupb6vn
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
98,294
Tissue Fixation | HubMAP | JHU-TMC
0
dx.doi.org/10.17504/protocols.io.3byl4929jgo5/v2
https://www.protocols.io/view/tissue-fixation-hubmap-jhu-tmc-db8w2rxe
Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Miklhail James, Sashank Reddy, Denis Wirtz
TITLE: Tissue Fixation | HubMAP | JHU-TMC AUTHORS: Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Miklhail James, Sashank Reddy, Denis Wirtz [DESCRIPTION] First and most important - the original tissue sample must be of good quality. Factors such as warm ischemic time, the time delay between tissue excisio...
["[Tissue Fixation] Tissue is harvested using our protocol (Tissue Harvesting Protocol)", "[Tissue Fixation] The rate of penetration of formaldehyde depends on the size of the biopsy. Trimming is often required to facilitate fixation. Tissues placed in the tissue cassettes should be no thicker than 3-4mm because the in...
70,241
Phage DNA extraction with Monarch kit and digestion to single nucleosides
1
dx.doi.org/10.17504/protocols.io.3byl4j2p8lo5/v1
https://www.protocols.io/view/phage-dna-extraction-with-monarch-kit-and-digestio-cgt9twr6
Atanas Radkov, Adair Borges
TITLE: Phage DNA extraction with Monarch kit and digestion to single nucleosides AUTHORS: Atanas Radkov, Adair Borges [DESCRIPTION] This protocol details high-molecular-weight DNA extraction from bacteriophages using the NEB Monarch HMW DNA Extraction Kit for Tissue. Following DNA extraction, DNA is digested down to...
["[Nucleoside digestion] Use a Nucleoside Digestion Mix (NEB - M0649S) to digest 1 µg of DNA. Each reaction should contain 1 µL of Nucleoside Digestion Mix and 2 µL of 10× Nucleoside Digestion Mix buffer.", "[Nucleoside digestion] Perform reactions in 20 µL total volume and incubate at 37 °C for 1 h, followed by enzyme...
39,154
Opti-Prep density separation of viruses for metagenomics
4
dx.doi.org/10.17504/protocols.io.261geobqwl47/v1
https://www.protocols.io/view/opti-prep-density-separation-of-viruses-for-metage-bigskbwe
Aditi K. Narayanan, Alon Philosof
TITLE: Opti-Prep density separation of viruses for metagenomics AUTHORS: Aditi K. Narayanan, Alon Philosof [DESCRIPTION] Density purification of viruses from liquid suspensions to remove cells, cell debris, and contaminants related to growth media. Purified viruses can be used in a variety of downstream applications, ...
["[Preparation of OptiPrep gradient] Prepare OptiPrep solutions at your chosen densities (OptiPrep Density Gradient Medium, Millipore Sigma, SKU:D1556). OptiPrep is a 60% w/v solution of iodixanol (density 1.32 g/mL). It comes sterile, with a rubber stopper in the top. Wipe the stopper with 70% ethanol and use a needle...
91,459
Purification of Methylglucose Lipopolysaccharides (MGLP)
6
dx.doi.org/10.17504/protocols.io.n92ldmjexl5b/v1
https://www.protocols.io/view/purification-of-methylglucose-lipopolysaccharides-c5jby4in
Briana Marisco, Luisa M Nieto Ramirez, Karen Dobos
TITLE: Purification of Methylglucose Lipopolysaccharides (MGLP) AUTHORS: Briana Marisco, Luisa M Nieto Ramirez, Karen Dobos [DESCRIPTION] Methylglucose lipopolysaccharides (mGLPs) are poly-methylated polysaccharides (PMPSs) that are acylated with short-chain fatty acids, uniquely synthesized by bacteria belonging to t...
["[Total lipid extraction] Obtain approximately 100 g Mycobacterium tuberculosis (Mtb) cells and freeze dry by lyophilization.", "[Total lipid extraction] Determine the dry weight of the cells and transfer them to a 500 mL glass bottle.", "[Total lipid extraction] Delipidate cells by adding 10mL of 10:10:3 [vol/vol/v...
11,434
ROS staining
null
dx.doi.org/10.17504/protocols.io.peidjce
null
Magdalena Julkowska
TITLE: ROS staining AUTHORS: Magdalena Julkowska [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol adapted from Shin et al., 2005 and Orman-Ligeza et al. 2016 (citing Shin)</div></div> [STEPS] ?. [Transfer the seedlings from agar plates into nutrient solution containing DCF-DA (50 uM) for 30...
["[Transfer the seedlings from agar plates into nutrient solution containing DCF-DA (50 uM) for 30 min. Preferably in darkness as DCF-DA is light sensitive]", "[wash with medium without DCF-DA and image fluorescence microscopy (460–500 nm bandpass excitation. Emission of 510– 560 nm bandpass]", "[wash with medium witho...
93,770
Preparation of Recording ACSF (artificial cerebrospinal fluid) for slice electrophysiology
4
dx.doi.org/10.17504/protocols.io.x54v9p4nmg3e/v1
https://www.protocols.io/view/preparation-of-recording-acsf-artificial-cerebrosp-c7tiznke
Sarthak M. Singhal, thnasko
TITLE: Preparation of Recording ACSF (artificial cerebrospinal fluid) for slice electrophysiology AUTHORS: Sarthak M. Singhal, thnasko [DESCRIPTION] Hnasko lab- preparation of Recording ACSF Recording ACSF is used for incubating brain slices during cell-attached or whole-cell electrophysiological recordings of neurons...
["Prepare 10X stock solution.", "Composition of 1 L of 10X Recording ACSF (in mM): 1250 NaCl, 25 KCl, 12 NaH2PO4, 125 d-glucose, 20 CaCl2 and 20 MgSO4.", "Composition of 1 L of 10X Recording ACSF (in g): 73.05 NaCl, 1.86 KCl, 1.44 NaH2PO4, 22.52 d-glucose, 2.94 CaCl2 and 4.93 MgSO4.", "Dissolve all salts in 1 L of ddH2...
43,250
V.3 - Direct wastewater RNA capture and purification via the "Sewage, Salt, Silica and SARS-CoV-2 (4S)" method
4
dx.doi.org/10.17504/protocols.io.bngsmbwe
https://www.protocols.io/view/v-3-direct-wastewater-rna-capture-and-purification-bngsmbwe
Oscar Whitney, Basem Al-Shayeb, Alex Crits-Cristoph, Mira Chaplin, Vinson Fan, Hannah Greenwald, Adrian Hinkle, Rose Kantor, Lauren Kennedy, Anna Maurer, Robert Tjian, Kara L. Nelson, UC Berkeley Wastewater-based epidemiology consortium
TITLE: V.3 - Direct wastewater RNA capture and purification via the "Sewage, Salt, Silica and SARS-CoV-2 (4S)" method AUTHORS: Oscar Whitney, Basem Al-Shayeb, Alex Crits-Cristoph, Mira Chaplin, Vinson Fan, Hannah Greenwald, Adrian Hinkle, Rose Kantor, Lauren Kennedy, Anna Maurer, Robert Tjian, Kara L. Nelson, UC Berkel...
["[Preparing RNA wash buffers]\nPrepareeach of two wash buffers - Wash buffer #1 (4S-WB1) and #2 (4S-WB2), for later use during cleanup of RNA bound to silica columns.\n1 L", "[Preparing RNA wash buffers]\n4S-WB1 composition: ABCD1ReagentOriginal molarity/%Final molarity/%Volume per liter of buffer2NaCl5 M1.5 M300 mL...
null
null
null
dx.doi.org/10.17504/protocols.io.dmd425
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <strong>Materials:</strong> <br />- 10 place filter manifold<br /> - Towers (chilled overnight @ 4C) <br />- 5% (w/v) Trichloroacetic acid (TCA) (chilled overnight @ 4C) Usually 1L is enough for a single time series set <br />- 32 15ml centrifuge tubes <br />- Pump <br />- Scinti...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.p9ydr7w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div title="Page 4"> <div> <div> <p><span style="font-size: 9.000000pt; font-family: 'AdvHelv_R'; color: rgb(13.725000%, 12.157000%, 12.549000%);">Experimental set up for BONCAT coupled with fluorophore addition via copper (I)- catalysed azide-alkyne cycloaddition (i.e., click c...
[]
97,460
Drosophila melanogaster inoculation with Bd
0
dx.doi.org/10.17504/protocols.io.yxmvme7n6g3p/v1
https://www.protocols.io/view/drosophila-melanogaster-inoculation-with-bd-dbeu2jew
Tatum S Katz, Alyssa Byer, Renwei Chen
TITLE: Drosophila melanogaster inoculation with Bd AUTHORS: Tatum S Katz, Alyssa Byer, Renwei Chen [DESCRIPTION] Methods to inoculate D. melanogaster with Batrachochytrium dendrobatidis. For manuscript Drosophila melanogaster is a possible vector of Batrachochytrium dendrobatidis under review at PLOS One. [STEPS] S...
["[Protocol] Plate 0.5ml of treatment (10^6 zsps per fly or DI water) onto each agar plate", "Allow plates to dry so that no water is puddled on the surface", "Place 10 flies onto each plate", "Seal plates with parafilm and incubate at room temperature for 24 hours", "Transfer flies into fly vials (keeping all flies fr...
61,519
An optimized protocol for flow virometry (FVM)-based detection and enumeration of T4 bacteriophage
1
null
https://www.protocols.io/view/an-optimized-protocol-for-flow-virometry-fvm-based-b8bprsmn
Hannah Safford
TITLE: An optimized protocol for flow virometry (FVM)-based detection and enumeration of T4 bacteriophage AUTHORS: Hannah Safford [DESCRIPTION] Flow virometry can support advanced water treatment and reuse by delivering near real-time information about viral water quality. Realizing the full potential of this techni...
["[Preparation of purified, high-titer T4 stock] Propagate T4 and host from freeze-dried specimens.", "[Preparation of purified, high-titer T4 stock] Use propagated T4 and host to prepare purified, high-titer T4 stock for FVM.", "[Preparation of purified, high-titer T4 stock] Aliquot propagated specimens and store at -...
96,343
bHEV genotpying RT-PCR
0
null
https://www.protocols.io/view/bhev-genotpying-rt-pcr-dabx2apn
Siu Fung Ho
TITLE: bHEV genotpying RT-PCR AUTHORS: Siu Fung Ho [DESCRIPTION] bHEV genotpying RT-PCR [STEPS] SECTION: Polyermase Chain Reaction 14. Setup mastermix in a 1.5 mL eppendorf tube. The following amounts are for a single reaction Component Volume 2x Reaction Mix 25 µL For...
["[Polyermase Chain Reaction] Setup mastermix in a 1.5 mL eppendorf tube. The following amounts are for a single reaction\n\nComponent Volume\n2x Reaction Mix 25 µL \nForward Primer 10 μM 2 µL\nReverse Primer 10 μM 2 µL\nNuclease-free water 16 µL \nTotal ...
27,394
Loading Tn5 Enzyme with sci-protocol Oligonucleotides
1
dx.doi.org/10.17504/protocols.io.6zahf2e
https://www.protocols.io/view/loading-tn5-enzyme-with-sci-protocol-oligonucleoti-6zahf2e
Ryan Mulqueen, Andy Fields, Andrew Adey
TITLE: Loading Tn5 Enzyme with sci-protocol Oligonucleotides AUTHORS: Ryan Mulqueen, Andy Fields, Andrew Adey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Loading of Tn5 enzyme for sci-ATAC and s3 protocols.</div></div> [STEPS] ?. [Prepare Reagents] Prepare for protein dilution. ABCD1...
["[Prepare Reagents]\nPrepare for protein dilution. ABCD1ReagentStock ConcentrationFinal ConcentrationAmount of Stock2HEPES-KOH (pH 7.2)1M100mM5mL3NaCl5M200mM2mL4Glycerol100%25%12.5mL5Triton-X100100%0.2%100uL6ddH2O30.4mL (to 50mL)7DTTDry2mM15.4 mg\n[2.125X Tn5 Dilution buffer]\nABCD1ReagentStock ConcentrationFinal ...
47,004
First-strand synthesis and touchdown PCR for SARS-CoV-2
3
dx.doi.org/10.17504/protocols.io.br54m88w
https://www.protocols.io/view/first-strand-synthesis-and-touchdown-pcr-for-sars-br54m88w
Leonardo Caserta
TITLE: First-strand synthesis and touchdown PCR for SARS-CoV-2 AUTHORS: Leonardo Caserta [STEPS]
[]
50,944
Human endometrium and endometriosis tissue dissociation for single-cell RNA sequencing
4
dx.doi.org/10.17504/protocols.io.bvy8n7zw
https://www.protocols.io/view/human-endometrium-and-endometriosis-tissue-dissoci-bvy8n7zw
Yuliana Tan, Diane Luo, Suleyman Bozal, Paul Robson, Elise Courtois
TITLE: Human endometrium and endometriosis tissue dissociation for single-cell RNA sequencing AUTHORS: Yuliana Tan, Diane Luo, Suleyman Bozal, Paul Robson, Elise Courtois [DESCRIPTION] Tissue dissociation is critical step to ensure the quality of samples for single-cell RNA sequencing. To ensure recovery of most cells...
["[Buffer and Enzymes Preparation] Prepare fresh digestion enzyme - Cold Active Protease (CAP) solution:", "[Viable Cell Sorting] Stain cells with viability dye (Propidium Iodide and Calcein Violet) following manufacturer's protocol, incubate 20 min.", "[Tissue Digestion with CAP] Mince the tissue to 1 mm2 cube with st...
51,860
Orange peel | Chitosan | Hemp
1
null
https://www.protocols.io/view/orange-peel-chitosan-hemp-bwvupe6w
Jil A Berenblum Phoenix Lai Esther Chang Anna Lu
TITLE: Orange peel | Chitosan | Hemp AUTHORS: Jil A Berenblum Phoenix Lai Esther Chang Anna Lu [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Orange peel | Chitosan | Hemp</span></div></div> [STEPS] ?. Remove the peel from an orange(we used Citrus Sinensis N...
["Remove the peel from an orange(we used Citrus Sinensis Navel Oranges from Florida). While the peel is still fresh, blend it into small granules. The size of the granules can affect the characteristics of the end material.", "In a bowl, combine the Hemp Protein Powder , Gluten , and granulated Orange Peel Mix the in...
17,598
DNA extraction from avian faeces stored in ethanol
1
dx.doi.org/10.17504/protocols.io.ve6e3he
https://www.protocols.io/view/dna-extraction-from-avian-faeces-stored-in-ethanol-ve6e3he
James Nicholls
TITLE: DNA extraction from avian faeces stored in ethanol AUTHORS: James Nicholls [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">DNA extraction from blue tit (Cyanistes caeruleus) faeces stored in ethanol.</span></div><div class = "text-block"><span>Methodology use...
["Remove ~100-200 mg of blue tit faeces (typically 2-3 small fragments of faeces, each ~5 mm long) from storage tube, allow ethanol to evaporate off, and then place into a 2 mL round-bottomed centrifuge tube (Eppendorf 2 mL SafeLock tubes are good). Avoid using faeces that has lots of uric acid on it, or scrape off ur...
null
null
null
dx.doi.org/10.17504/protocols.io.efrbbm6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol provides a method to predict phage community diversity using the algorithm PHACCS. Sequencing of viral communities often results in a high percent of unknown reads, largely due to our incomplete reference databases. To address this unknown factor of viromes, the al...
[]
92,680
FAST.R Installation and Launch with RStudio
4
dx.doi.org/10.17504/protocols.io.kxygx3464g8j/v1
https://www.protocols.io/view/fast-r-installation-and-launch-with-rstudio-c6rgzd3w
Francesco Neri
TITLE: FAST.R Installation and Launch with RStudio AUTHORS: Francesco Neri [DESCRIPTION] Step -by-step protocol to setup the R shiny application FAST.R for data analysis and visualization of FAST-generated data. This protocol is meant to be used within the FAST workflow [STEPS] SECTION: Install FAST.R with RStudio 1....
["[Install FAST.R with RStudio] If familiar with R and RStudio, see the FAST.R package page for installation and use.\n\nIf NOT familiar with R and RStudio, read instructions below", "[Install FAST.R with RStudio] Install FAST-R", "[Install FAST.R with RStudio] Download and install R and RStudio\nDownload from here. No...
24,457
Transformation of the chlorarachniophyte Amorphochlora amoebiformis by electroporation
null
dx.doi.org/10.17504/protocols.io.35hgq36
null
Kodai Fukuba, Liz Cooney, Nick Irwin, Patrick J. Keeling, Yoshihisa Hirakawa
TITLE: Transformation of the chlorarachniophyte Amorphochlora amoebiformis by electroporation AUTHORS: Kodai Fukuba, Liz Cooney, Nick Irwin, Patrick J. Keeling, Yoshihisa Hirakawa [STEPS] ?. [Plasmid preparation] Propagate plasmid DNA (pLaRGfp or its derivatives, such as pLaR182+GFP) in the Escherichia coli strain DH5...
["[Plasmid preparation]\nPropagate plasmid DNA (pLaRGfp or its derivatives, such as pLaR182+GFP) in the Escherichia coli strain DH5α.", "[Plasmid preparation]\nPurify plasmid DNA from 200 mL culture of E. coli by a Qiagen Plasmid Maxi Kit (Qiagen).", "[Plasmid preparation]\nAdjust plasmid DNA concentration to 3-5 µg/µL...
18,598
FCA general dissociation protocol
null
dx.doi.org/10.17504/protocols.io.weefbbe
null
JONGEUN PARK
TITLE: FCA general dissociation protocol AUTHORS: JONGEUN PARK [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is the protocol used for FCA_ImmunoP project</div></div> [STEPS]
[]
13,469
RAD barcodes
3
null
https://www.protocols.io/view/rad-barcodes-rd5d286
Tomasz Suchan
TITLE: RAD barcodes AUTHORS: Tomasz Suchan [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">File listing 96 barcodes on SbfI-compatible P1 adapters we use for RAD-seq libraries.</div></div> [STEPS]
[]
33,575
Plant and fungal DNA extraction using a QIAGEN QIAxtractor
null
dx.doi.org/10.17504/protocols.io.bc2fiybn
null
Laura Forrest, Michelle Hart, Alexandra Clark
TITLE: Plant and fungal DNA extraction using a QIAGEN QIAxtractor AUTHORS: Laura Forrest, Michelle Hart, Alexandra Clark [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Here we describe a semi-robotic plate DNA extraction method that we have used at the Royal Botanic Garden Edinburgh (RBGE) to proce...
["Lysis pre-treatment: in a trough mix 4 μl DX enzyme (kept at 4° C in the fridge) and 416 μl Tissue Digest (DXT) per sample (master mix 40.4 ml DXT and 392 μl DX enzyme per 96-sample plate); if extracting from fresh tissue rather than silica-dried or herbarium samples, substitute 4 μl DXT with 4 μl RNase A.", "Separat...
69,719
Multiplex fluorescent immunostaining of thin, fixed mouse brain tissue sections to characterize human iPSC-derived cell xenografts
1
dx.doi.org/10.17504/protocols.io.4r3l275mxg1y/v1
https://www.protocols.io/view/multiplex-fluorescent-immunostaining-of-thin-fixed-cgbxtspn
Benjamin Trist, Louise Cottle
TITLE: Multiplex fluorescent immunostaining of thin, fixed mouse brain tissue sections to characterize human iPSC-derived cell xenografts AUTHORS: Benjamin Trist, Louise Cottle [DESCRIPTION] This protocol describes our multiplex fluorescent immunohistochemistry protocol used to identify human iPSC-derived cells within...
["[Day 1 (~3-4 hrs)] Pre-heat oven and Antigen Retrieval (AR) buffer to 70 °C.", "[Day 1 (~3-4 hrs)] Label scintillation vials to match labels on section storage plates (mouse and section series IDs, name, date etc.).", "[Day 1 (~3-4 hrs)] Transfer sections into scintillation vials using a transfer pipette or fine pain...
58,649
Processing of Radial K-space DW-MRI Data
1
dx.doi.org/10.17504/protocols.io.j8nlkkwj1l5r/v1
https://www.protocols.io/view/processing-of-radial-k-space-dw-mri-data-b5hzq376
Miguelrj , Hee Kwon Song, Stephen Pickup, Mamtaaryagupta , Rong Zhou
TITLE: Processing of Radial K-space DW-MRI Data AUTHORS: Miguelrj , Hee Kwon Song, Stephen Pickup, Mamtaaryagupta , Rong Zhou [DESCRIPTION] The protocol includes reconstruction of diffusion weighted images from radial k-space data and using a 3-parameter fit to derive metrics of ADC and kurtosis index. [STEPS] 1...
["Image reconstruction\n\nRadially acquired diffusion-weighted images (DWIs) are reconstructed using the following steps:\n\nApply zero-order phase correction to each radial spoke using the average phase offset at the center slice of the lowest b-value image\nZerofill k-space by a factor of 2 to double field of view\nM...
68,823
RNA sequencing analysis of transcriptomic responses to vagal nerve stimulation in myenteric ganglia of porcine colon
1
dx.doi.org/10.17504/protocols.io.eq2lynmeqvx9/v1
https://www.protocols.io/view/rna-sequencing-analysis-of-transcriptomic-response-cffxtjpn
Tao Li, Pu-Qing Yuan
TITLE: RNA sequencing analysis of transcriptomic responses to vagal nerve stimulation in myenteric ganglia of porcine colon AUTHORS: Tao Li, Pu-Qing Yuan [DESCRIPTION] The region-dependent molecular characterization and influences by vagus nerve stimulation (VNS) influences in the porcine ENS has been little investiga...
["Six castrated male Yucatan minipigs (3 of each controls and 3 with VNS) were fasted for 12 h and anaesthetized. Those with VNS underwent electrical stimulation of the celiac branch of the abdominal vagus nerve (2 Hz, 0.3-4 ms, 5 mA, 10 min) using pulse train.", "The proximal and distal colonic (pC, dC) pecimens with ...
54,149
Ligation and gel electrophoresis
4
dx.doi.org/10.17504/protocols.io.by5dpy26
https://www.protocols.io/view/ligation-and-gel-electrophoresis-by5dpy26
Chia-Hsien Shih
TITLE: Ligation and gel electrophoresis AUTHORS: Chia-Hsien Shih [DESCRIPTION] This protocol is used for ligation of circular probe [STEPS] SECTION: Preparation 1. Dilute linear DNA into 10µM SECTION: Preparation 2. Dilute T4 ligation primer into 10µM SECTION: Protocol of Ligation 3. Add 11 µL of RNase-free water i...
["[Preparation] Dilute linear DNA into 10µM", "[Preparation] Dilute T4 ligation primer into 10µM", "[Protocol of Ligation] Add 11 µL of RNase-free water into a eppendorf", "[Protocol of Ligation] Add 3 µL of 10X T4 Ligase buffer", "[Protocol of Ligation] Add 9 µL of 10µM T4 ligation primer", "[Protocol of Ligation] Add...
84,050
Cortical, Striatal and Dopaminergic Neurons Cultures Protocol
1
dx.doi.org/10.17504/protocols.io.rm7vzxn48gx1/v1
https://www.protocols.click/view/cortical-striatal-and-dopaminergic-neurons-culture-cwbsxane
giulia.tombesi
TITLE: Cortical, Striatal and Dopaminergic Neurons Cultures Protocol AUTHORS: giulia.tombesi [DESCRIPTION] This protocol describes how to obtain primary neuronal cultures from postnatal mice (P0-P1). It can be used to obtain pure cortical neurons cultures or stratal-cortical neurons and dopaminergic-cortical neurons c...
["[Cells Dissociation] Snip the tissues into smaller pieces and place them in Papain solution. Use around 500 µL of Papain solution for 5 SN, around 1 mL of Papain solution for 5 striata and around 5 mL of Papain solution for 5-6 cortices.", "[Cells Dissociation] Triturate 10 times with a 5ml-pipette", "[Cells Dissocia...
null
null
null
dx.doi.org/10.17504/protocols.io.t5teq6n
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
87,084
Post-diagnosis dietary patterns among cancer survivors in relation to all-cause mortality and cancer-specific mortality: a systematic review and meta-analysis of cohort studies
1
dx.doi.org/10.17504/protocols.io.yxmvm3en5l3p/v1
https://www.protocols.io/view/post-diagnosis-dietary-patterns-among-cancer-survi-czakx2cw
Maria-Eleni Spei, Ioannis Bellos, esamoli, Vassiliki Benetou
TITLE: Post-diagnosis dietary patterns among cancer survivors in relation to all-cause mortality and cancer-specific mortality: a systematic review and meta-analysis of cohort studies AUTHORS: Maria-Eleni Spei, Ioannis Bellos, esamoli, Vassiliki Benetou [DESCRIPTION] This study aims to synthesize the latest evidence r...
["Objective: To synthesize the latest evidence regarding the association of a priori and\na posteriori dietary patterns with robust outcome measures, such as total mortality and cancer-specific mortality.", "Eligibility criteria: Studies will be eligible to be included if: (i) they have a cohort design, prospective\nor...
45,982
C-SOP-401: Quality Control (QC) of DNA Libraries for Whole Genome Sequencing
4
null
https://www.protocols.io/view/c-sop-401-quality-control-qc-of-dna-libraries-for-bq56my9e
Mihir Kekre
TITLE: C-SOP-401: Quality Control (QC) of DNA Libraries for Whole Genome Sequencing AUTHORS: Mihir Kekre [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">The success of fragmentation and size selection is best confirmed using an electrophoretic instrum...
["[Before Starting]\nPrior to initiating the protocol, ensure that all active workbenches are cleaned with 80% ethanol, all relevant personal protective clothing is worn and the work area is prepared according to local GLP guidelines for molecular methods.Create an organised bench space by clearing away all clutter in ...
null
null
null
dx.doi.org/10.17504/protocols.io.k92cz8e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>Intended use:</strong></p> <p>Kinetic system for Aspartate Aminotransferase (AST) or Glutamic oxalacetic trasminase (GOT) determination.</p> <p><strong>Teste Principle:</strong></p> <p><strong>AST</strong> catalyzes specifically  the transfer of amino groups of asparc...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.njbdcin
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>Objective </strong></p> <p>To provide an in-depth and comprehensive overview of tick-borne rickettsioses among humans, livestock, wild animals and arthropods in Iran, a systematic review and meta-analysis will be performed. This review will capture the existing data r...
[]
69,136
Wet Chemistry RNA Isolation Method for Mature Cassava Leaf Tissue
1
null
https://www.protocols.io/view/wet-chemistry-rna-isolation-method-for-mature-cass-cfrqtm5w
Lynn Doran
TITLE: Wet Chemistry RNA Isolation Method for Mature Cassava Leaf Tissue AUTHORS: Lynn Doran [DESCRIPTION] RNA extraction protocol for cassava for RNA of sufficient quality and quantity to perform RT-PCR gene expression studies. *Low recovery was achieved using Qiagen RNeasy kit. Typical recoveries using the mini...
[]
46,604
Genotyping of polymorphisms rs1801133 in MTHFR, rs1051266 in SLC19A1 and rs1805087 in MTR
1
dx.doi.org/10.17504/protocols.io.brrkm54w
https://www.protocols.io/view/genotyping-of-polymorphisms-rs1801133-in-mthfr-rs1-brrkm54w
Nayane Soares de Lima, Caroline Christine Pincela da Costa , Rômulo Morais Azevedo, Angela Adamski Reis, Rodrigo da Silva Santos
TITLE: Genotyping of polymorphisms rs1801133 in MTHFR, rs1051266 in SLC19A1 and rs1805087 in MTR AUTHORS: Nayane Soares de Lima, Caroline Christine Pincela da Costa , Rômulo Morais Azevedo, Angela Adamski Reis, Rodrigo da Silva Santos [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div clas...
["[DNA extraction]\n.justify:after {\n\t\t\t\t\t\t\t content: \"\";\n\t\t\t\t\t\t\t display:inline-block;\n\t\t\t\t\t\t\t width: 100%;\n\t\t\t\t\t\t\t }\n\t\t\t\t\t\t\tThe DNA samples were extracted from peripheral blood and submitted to DNA purification using the commercial extraction kit, following the prot...
50,301
Study Medications (Part 5 of Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-onset Type 1 Diabetes Mellitus)
1
dx.doi.org/10.17504/protocols.io.bvc5n2y6
https://www.protocols.io/view/study-medications-part-5-of-safety-and-efficacy-of-bvc5n2y6
Stephen.Gitelman , Jeffrey A. Bluestone
TITLE: Study Medications (Part 5 of Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-onset Type 1 Diabetes Mellitus) AUTHORS: Stephen.Gitelman , Jeffrey A. Bluestone [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is Part 5 of "Safety and Efficacy of Imatinib for Preserv...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.gwubxew
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol for cell separation, enumeration, and sorting of low biomass samples for SIP-NanoSIMS preparations. </p> [BEFORE_START] <ol> <li>Clean 12 spoons <ol> <li>soaked in bleach ~15min, then rinse with 0.2um MQ, then Sonicate 15 min, then rinse, then bring water cup with s...
[]
45,426
Modified pH drop protocol (K+)
4
null
https://www.protocols.io/view/modified-ph-drop-protocol-k-bqksmuwe
Elizabeth Fozo
TITLE: Modified pH drop protocol (K+) AUTHORS: Elizabeth Fozo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Modified pH drop protocol (K</span><span style = "vertical-align:super;">+</span><span style = "vertical-align:super;">)</span></div></div> [STEPS] ?. [To prepare the K+ electrode for...
["[To prepare the K+ electrode for use:]\nRemove the rubber caps.", "[Do slope check for electrode performance:]", "[Standard curve:]\n0mM (no K+) = 9mL water + 1mL 1M NaCl", "[Modified pH drop protocol:]\nRead pH & mV readings atdesired intervals.", "[To prepare the K+ electrode for use:]\nFill with 0.1M NaCl filling ...
82,336
shRNA plasmids
3
dx.doi.org/10.17504/protocols.io.yxmvm288bg3p/v1
https://www.protocols.io/view/shrna-plasmids-cum8wu9w
Shiyi Wang
TITLE: shRNA plasmids AUTHORS: Shiyi Wang [DESCRIPTION] How to make shRNA plasmids [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.cpvvn5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Basic Protocol for culturing Neuro2a cells. [GUIDELINES] <strong>The following are taken directly from the vendor website, I use high glucose DMEM instead of Eagle's MEM. </strong><br /><br /><strong>Complete Growth Medium</strong> The base medium for this cell line is ATCC-for...
[]
58,212
Gallyas-silver stain
4
dx.doi.org/10.17504/protocols.io.b44cqysw
https://www.protocols.io/view/gallyas-silver-stain-b44cqysw
Christiana.bjorkli , María José Lagartos Donate
TITLE: Gallyas-silver stain AUTHORS: Christiana.bjorkli , María José Lagartos Donate [DESCRIPTION] Simplified Gallyas-silver staining protocol that works for pathological staining in rodent and human tissue [STEPS] SECTION: Solutions that can be made in advance and stored in a fume hood 2. SECTION: Solutions that ...
["[Solutions that can be made in advance and stored in a fume hood]", "[Solutions that can be made in advance and stored in a fume hood] Make 4% of NaOH (sodium hydroxide) - use gloves, store in fume hood", "[Solutions that can be made in advance and stored in a fume hood]", "[Solutions that can be made in advance and ...
36,053
HuBMAP UF TMC - Single Cell RNA Seq (10x Genomics) Library Preparation and Sequencing 
null
dx.doi.org/10.17504/protocols.io.bffvjjn6
https://www.protocols.io/view/hubmap-uf-tmc-single-cell-rna-seq-10x-genomics-lib-bffvjjn6
Franchesca Farris, Maigan Brusko
TITLE: HuBMAP UF TMC - Single Cell RNA Seq (10x Genomics) Library Preparation and Sequencing  AUTHORS: Franchesca Farris, Maigan Brusko [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for the generation of 3' (chemistry version 3) cDNA libraries from individual cells via droplet gen...
["[GEM Generation & Barcoding]\nPreparation of Single Cell Master Mix: a. Prepare Master Mix on ice. Pipette mix 15x and centrifuge briefly. b. Add 33.4 µl Master Mix into each tube of a PCR 8-tube strip on ice.", "[GEM Generation & Barcoding]\nLoad Chromium Chip B: a. Assemble Chromium Chip B in a 10x Chip H...
null
null
null
dx.doi.org/10.17504/protocols.io.etpbemn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This tutorial was based off of the paper The microbes we eat: abundance and taxonomy of microbes consumed in a day's worth of meals for three diet types. <span style="font-weight: 400;">The article provides those interested in bioinformatics, food science, or microbiology inform...
[]
97,509
CODA (part 3): deep learning tissue structures labeling | HuBMAP | JHU-TMC
0
dx.doi.org/10.17504/protocols.io.81wgbz1x3gpk/v1
https://www.protocols.io/view/coda-part-3-deep-learning-tissue-structures-labeli-dbgd2js6
Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Sashank Reddy, Denis Wirtz
TITLE: CODA (part 3): deep learning tissue structures labeling | HuBMAP | JHU-TMC AUTHORS: Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Sashank Reddy, Denis Wirtz [DESCRIPTION] In this section, we describe steps to create a basic semantic segmentation algorithm using CODA. CODA uses a modified resnet50 net...
["[Deep learning multi-labelling of tissue structures using training on manual annotations] Choose the biological structures you wish to segment in your images. Semantic segmentation algorithms must classify every pixel of every image with a label, so your list must be exhaustive. For example, in lung histology you cou...
68,786
ES cells general methods
4
dx.doi.org/10.17504/protocols.io.e6nvwkxz9vmk/v1
https://www.protocols.io/view/es-cells-general-methods-cfestjee
Rebecca Berrens
TITLE: ES cells general methods AUTHORS: Rebecca Berrens [DESCRIPTION] general methods for mouse ES cell culturing [BEFORE_START] All media should be warmed up to37 °C in a water-bath before use. Spray all the bottles (except new culture flasks or tubes) before putting into the hood. [STEPS] SECTION: Reactivating ce...
["[Reactivating cells from liquid nitrogen stock] Spray a tube with 70% EtOH and wipe off. Transfer all thawed cells (~1ml) into the 15ml Falcon tube.", "[Reactivating cells from liquid nitrogen stock] Drop by drop, and swirl from time to time, add 9ml ES complete medium or EC10 medium. Final volume of cells is 10ml. (...
82,571
Immunohistochemistry for FOXP3+ staining in Breast Cancer Tissue
6
dx.doi.org/10.17504/protocols.io.eq2ly7m6elx9/v1
https://www.protocols.click/view/immunohistochemistry-for-foxp3-staining-in-breast-cuvjww4n
Freda Halim
TITLE: Immunohistochemistry for FOXP3+ staining in Breast Cancer Tissue AUTHORS: Freda Halim [DESCRIPTION] These are protocols used for study of FOXP3+ Cell Count in Luminal B Her-2 negative BC patients. We used using paraffin sections of 66 samples and stained the slides for FOXP3+ antibody, using primary antibody cl...
["[Deparaffinization and Rehydration] Incubate slides in Xylenes for 3 minutes", "[Deparaffinization and Rehydration] Incubate slides in Xylenes", "[Deparaffinization and Rehydration] Incubate slides in Xylenes", "[Deparaffinization and Rehydration] Rehydrate slides in 100% Ethanol", "[Deparaffinization and Rehydration...
33,146
LCMS-SOP Determination of Microcystins in Water Samples by High Performance Liquid Chromatography (HPLC) with Single Quadrupole Mass Spectrometry (MS)
3
dx.doi.org/10.17504/protocols.io.bck2iuye
https://www.protocols.io/view/lcms-sop-determination-of-microcystins-in-water-sa-bck2iuye
Gregory L Boyer
TITLE: LCMS-SOP Determination of Microcystins in Water Samples by High Performance Liquid Chromatography (HPLC) with Single Quadrupole Mass Spectrometry (MS) AUTHORS: Gregory L Boyer [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This standard operating procedure is used to determine and quantify m...
[]
58,949
Overall protocol for top-down CZE-MS/MS of human small intestine tissue
1
dx.doi.org/10.17504/protocols.io.b5tdq6i6
https://www.protocols.io/view/overall-protocol-for-top-down-cze-ms-ms-of-human-s-b5tdq6i6
Jeannie Camarillo, Bryon Drown, Neil Kelleher
TITLE: Overall protocol for top-down CZE-MS/MS of human small intestine tissue AUTHORS: Jeannie Camarillo, Bryon Drown, Neil Kelleher [DESCRIPTION] Overview description of the process for acquiring top-down capillary zone electrophoresis MS/MS data on human small intestine tissue [STEPS] SECTION: Tissue Collection...
["[Tissue Collection] Small intestine tissue was provided by Stanford University TMC. Tissue was collected according to the following: \n\nhttps://dx.doi.org/10.17504/protocols.io.bin8kdhw", "[Sample Preparation]", "[Data Acquisition]", "[Data Analysis]"]
78,515
Carbon nanotube (CNT) yarn fabrication for chronic interfacing with the autonomic nervous system
1
dx.doi.org/10.17504/protocols.io.n2bvj8enpgk5/v1
https://www.protocols.io/view/carbon-nanotube-cnt-yarn-fabrication-for-chronic-i-cqwtvxen
Grant Mccallum, Xiaohong Sui, Chen Qiu, Joseph Marmerstein, Yang Zheng, Thomas Eggers, Chuangang Hu, Liming Dai, Dominique Durand
TITLE: Carbon nanotube (CNT) yarn fabrication for chronic interfacing with the autonomic nervous system AUTHORS: Grant Mccallum, Xiaohong Sui, Chen Qiu, Joseph Marmerstein, Yang Zheng, Thomas Eggers, Chuangang Hu, Liming Dai, Dominique Durand [DESCRIPTION] The ability to reliably and safely communicate chronically wit...
["[Carbon nanotube (CNT) yarn fabrication] VA-MWCNT arrays (with MWCNTs of ca.250 μm in length andca.30 nm in diameter) (Fig. 1a), were first synthesized on Fe/Al-coated Si/SiO2 wafers using a low pressure chemical vapor deposition (CVD) method according to our previously reported procedure52.", "[Carbon nanotube (CNT)...
25,006
NEBNext® Ultra™ II End Repair/dA-Tailing Module (NEB #E7546)
null
dx.doi.org/10.17504/protocols.io.4nngvde
null
Menna Teffera, New England Biolabs
TITLE: NEBNext® Ultra™ II End Repair/dA-Tailing Module (NEB #E7546) AUTHORS: Menna Teffera, New England Biolabs [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The NEBNext Ultra II End Repair/dA-Tailing Module is optimized to convert 500 pg-1 μg of fragmented DNA to repaired DNA having 5 ́ phosphor...
["[NEBNext End Prep]\nMix the following contents in a sterile nuclease-free tube: AB1ComponentVolume2(green) NEBNext Ultra II End Prep Enzyme Mix3 µl3(green) NEBNext Ultra II End Prep Reaction Buffer7 µl4Fragmented DNA50 µl5Total Volume60 µl\nAB1ComponentVolume2(green) NEBNext Ultra II End Prep Enzyme Mix3 µl3(green) ...
null
null
null
dx.doi.org/10.17504/protocols.io.chut6v
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]