id
float64
1.55k
110k
title
stringlengths
1
256
template_id
float64
0
6
doi
stringlengths
39
49
url
stringlengths
40
92
authors
stringlengths
1
933
protocol_text
stringlengths
34
1.08M
steps_list
stringlengths
2
269k
67,190
Using the Thermo SPD1010 speedvac concentrator centrifuge for drying down peptides for LC-MS analysis
1
dx.doi.org/10.17504/protocols.io.36wgq78qyvk5/v1
https://www.protocols.io/view/using-the-thermo-spd1010-speedvac-concentrator-cen-cduws6xe
ronan.ocualain
TITLE: Using the Thermo SPD1010 speedvac concentrator centrifuge for drying down peptides for LC-MS analysis AUTHORS: ronan.ocualain [DESCRIPTION] This protocol details the procedure of using the Thermo SPD1010 speed vacuum concentrator centrifuge to dry peptide samples for storage and submission for LC-MS analysis. P...
["[Speedvac drying:] Turn the power switch located on the rear right hand side of the unit to the ON position. Wait 45 min before starting drying.", "[Speedvac drying:] After waiting for the system to come to temperature, add the vials to the centrifuge by placing the neck of the vial in and under the hole, and secure...
52,842
Day 3: Morphological Taxonomy
3
dx.doi.org/10.17504/protocols.io.bxuipnue
https://www.protocols.io/view/day-3-morphological-taxonomy-bxuipnue
Tom Little
TITLE: Day 3: Morphological Taxonomy AUTHORS: Tom Little [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Day 3: ADE practical instructions for morphological identifications</div></div> [STEPS]
[]
97,731
Paraffin embedding of tissue specimen | HuBMAP | JHU-TMC
0
dx.doi.org/10.17504/protocols.io.q26g71rn8gwz/v1
https://www.protocols.io/view/paraffin-embedding-of-tissue-specimen-hubmap-jhu-t-dbpb2min
Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Miklhail James, Sashank Reddy, Denis Wirtz
TITLE: Paraffin embedding of tissue specimen | HuBMAP | JHU-TMC AUTHORS: Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Miklhail James, Sashank Reddy, Denis Wirtz [DESCRIPTION] The standard for tissue embedding is to ensure tissue samples are appropriately oriented and embedded on the “cut edge” or flattest...
["[Orientation] The standard for tissue embedding is to ensure tissue samples are appropriately oriented and embedded on the “cut edge” or flattest edge available", "[Type of paraffin] the type of paraffin used in your laboratory should be optimal for your needs. Paraffin comes in many types. These types are differenti...
null
null
null
dx.doi.org/10.17504/protocols.io.fukbnuw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><span class="TextRun SCX24498567" lang="EN-US" style="font-size: 14pt; font-family: Times New Roman,serif; line-height: 23px;" xml:lang="EN-US"><span class="NormalTextRun SCX24498567" style="background-color: inherit;">This protocol is designed to allow you to clean large</sp...
[]
73,113
Luciferase Activity Assay
4
dx.doi.org/10.17504/protocols.io.dm6gpj37dgzp/v1
https://www.protocols.io/view/luciferase-activity-assay-cjmzuk76
Electra Brunialti, Alessandro Maria Villa, Paolo Ciana
TITLE: Luciferase Activity Assay AUTHORS: Electra Brunialti, Alessandro Maria Villa, Paolo Ciana [DESCRIPTION] Luciferase activity assay on cell extracts using Veritas (Turner) luminometer. [STEPS] 1. Lysate the cell using 1X Luciferase Cell Culture LysisReagent (Cat. E1531, Promega) following manufacturer instructio...
["Lysate the cell using 1X Luciferase Cell Culture LysisReagent (Cat. E1531, Promega) following manufacturer instruction;", "put 20 µl/well of cell lysate into the white plate (Cat 3912, Corning);", "prepare Reaction Buffer (100µl for each sample): 500 µM luciferin (Cat E160E, Promega), 20 mM Tricine, 0.1 mM EDTA, 1.07...
107,457
LRRK2 PhosphoSens Assay
0
dx.doi.org/10.17504/protocols.io.81wgbz8qngpk/v1
https://www.protocols.io/view/lrrk2-phosphosens-assay-dk694zh6
Nicolai D. Raig, Stefan Knapp
TITLE: LRRK2 PhosphoSens Assay AUTHORS: Nicolai D. Raig, Stefan Knapp [DESCRIPTION] With this enzymatic activity assay we were able to determine IC50 values of published inhibitors aswell as newly synthesized compounds that inhibit LRRK2. The assay is based on the in vitro phosphorylation reaction between the enzymati...
["Pipett a dilution series of eleven concentrations between 15µM and 0.4nM (calculated with an assay volume of 10µL) of the compounds into white 384-well plates (Greiner 781207) as duplicates with an ECHO acoustic dispenser (Labcyte). Pipett a equivalent of DMSO in two wells per compound as 0% (without protein and comp...
79,800
Tissue and blood collection
1
dx.doi.org/10.17504/protocols.io.bp2l69wmklqe/v1
https://www.protocols.click/view/tissue-and-blood-collection-cr6yv9fw
michela.deleidi, Bianca Marchetti, Federico Bertoli, Carmela Giachino
TITLE: Tissue and blood collection AUTHORS: michela.deleidi, Bianca Marchetti, Federico Bertoli, Carmela Giachino [DESCRIPTION] This protocol details tissue collection from mice. [STEPS] SECTION: Tissue and blood collection 1. For immunohistochemistry, mice (5-6 mice/genotype/age-group/treatment) were anesthetized an...
["[Tissue and blood collection] For immunohistochemistry, mice (5-6 mice/genotype/age-group/treatment) were anesthetized and transcardially perfused with 0.9% NaCl, followed by 4 % paraformaldehyde in phosphate buffer (pH 7.2 at 4°C), the brains and peripheral tissues carefully removed and post-fixed for 2-4 hrs, in 4%...
99,412
Food grade colorimetry of anthocyanins (A Youth Summer Camp Activity)
0
dx.doi.org/10.17504/protocols.io.yxmvmenqng3p/v1
https://www.protocols.io/view/food-grade-colorimetry-of-anthocyanins-a-youth-sum-ddbu22nw
Eric S McLamore, Geisianny AM Moreira, Diana Vanegas, dbahamo, Lisseth Casso-Hartmann, Lidadi Agbomi, Maria J Torres
TITLE: Food grade colorimetry of anthocyanins (A Youth Summer Camp Activity) AUTHORS: Eric S McLamore, Geisianny AM Moreira, Diana Vanegas, dbahamo, Lisseth Casso-Hartmann, Lidadi Agbomi, Maria J Torres [DESCRIPTION] This protocol describes activities used in summer education camps for youth (14 years and older). The ...
["[Anthocyanin activities for camp] Step 1) Gather and prepare materials\nGroup participants into teams of 2-4 people (Fig 1).\n\n \nTable 1 presents a planning table for a typical summer camp supporting 40 youth (ten teams of 4). For a camp of this size, 3-5 leaders are typically engaged to guide youth through activit...
null
null
null
dx.doi.org/10.17504/protocols.io.cp9vr5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Radioactive Labeling with&nbsp;T4 Polynucleotide Kinase 3' phosphatase minus [STEPS] ?. ?. ?.
[]
63,618
Measuring tension, pellet transit, and calcium imaging within cell subtypes in response to direct electrical field stimulation of colon
4
dx.doi.org/10.17504/protocols.io.36wgq7yokvk5/v1
https://www.protocols.io/view/measuring-tension-pellet-transit-and-calcium-imag-cadasa2e
Dante Heredia, Thomas Gould
TITLE: Measuring tension, pellet transit, and calcium imaging within cell subtypes in response to direct electrical field stimulation of colon AUTHORS: Dante Heredia, Thomas Gould [DESCRIPTION] Measurement of the effects of direct electrical stimulation of the mouse colon [BEFORE_START] Make sure there is enough g...
["A ventral midline incision is made and the entire colon is excised and placed into a sylguard-lined dissection dish filled with oxygenated Krebs-Ringers solution. The colon is then gently flushed of its contents. For all experiments, the colon is perfused with 35-degree, oxygenated Krebs-Ringers for the duration of ...
81,447
GFP Immunoprecipitation and Sample Preparation for Tandem Mass Tag (TMT) Mass Spectrometry Analysis
4
dx.doi.org/10.17504/protocols.io.eq2ly7kxqlx9/v1
https://www.protocols.io/view/gfp-immunoprecipitation-and-sample-preparation-for-ctsfwnbn
Prosenjit Pal, Raja S. Nirujogi, Francesca Tonelli, Dario R Alessi
TITLE: GFP Immunoprecipitation and Sample Preparation for Tandem Mass Tag (TMT) Mass Spectrometry Analysis AUTHORS: Prosenjit Pal, Raja S. Nirujogi, Francesca Tonelli, Dario R Alessi [DESCRIPTION] We describe a method to identify potential interactors of any Green Fluorescent Protein (GFP) tagged protein expressed in ...
["[Transient transfection of HEK293 cells] Plate cells in 10 cm dishes (one dish for each experimental condition) to give a 60-70% confluency the following day (around 2.2 x 106 cells seeded per 10 cm dish).", "[Transient transfection of HEK293 cells] Prepare a transfection mix in a sterile 1.5ml Eppendorf tube, contai...
86,474
Eastern Hemlock Tissue Collection for DNA
4
null
https://www.protocols.io/view/eastern-hemlock-tissue-collection-for-dna-cypixvke
Karl Fetter
TITLE: Eastern Hemlock Tissue Collection for DNA AUTHORS: Karl Fetter [DESCRIPTION] Steps for collecting tissue from Eastern Hemlocks for DNA analysis. The Plant Computational Genomics lab is conducting a landscape genomics study of climate adaptation in the Eastern Hemlock. This protocol is a step-by-step guide to co...
["[Introduction] This protocol is intended for field collection of leaf tissue for the Eastern Hemlock conservation genomics project sponsored by the Plant Computation Genomics lab at the University of Connecticut. The goal of the project is to identify climate adapted genomic variation for seed banking and potential u...
99,205
Lysosomal GCase (glucocerebrosidase) activity assay
0
dx.doi.org/10.17504/protocols.io.8epv5r9jdg1b/v1
https://www.protocols.io/view/lysosomal-gcase-glucocerebrosidase-activity-assay-dc5d2y26
Sara Gomes, Esther Sammler
TITLE: Lysosomal GCase (glucocerebrosidase) activity assay AUTHORS: Sara Gomes, Esther Sammler [DESCRIPTION] Here we report a method to measure enzyme activity of lysosomal glucocerebrosidase (GBA1, GCase) by monitoring the hydrolysis of the fluorescent substrate 4-methylumbelliferyl-β-D-glucopyranoside. The assay is ...
["[Buffer preparation] 0.1M citric acid: dissolve 19.2 g in 1 L", "[Buffer preparation] 0.2M sodium phosphate: dissolve 28.4 g in1 L.", "[Buffer preparation] Citrate-phosphate buffer, pH 5.4: mix 44.2 mLwith 56.8 mL to make 100mL citrate-phosphate buffer, pH 5.4.", "[Buffer preparation] 0.5M EDTA: dissolve 20.8 g in 8...
43,614
test 2
1
null
https://www.protocols.io/view/test-2-bnt6mere
Maria Guliakina
TITLE: test 2 AUTHORS: Maria Guliakina [STEPS] ?. test ?. ?.
["test"]
86,050
Unconventional secretion of alpha-synucein mediated by palmitoylated DNAJC5 oligomers
4
dx.doi.org/10.17504/protocols.io.rm7vzxb65gx1/v1
https://www.protocols.io/view/unconventional-secretion-of-alpha-synucein-mediate-cyaaxsae
iona.thomas-wright, Richard Wade-Martins
TITLE: Unconventional secretion of alpha-synucein mediated by palmitoylated DNAJC5 oligomers AUTHORS: iona.thomas-wright, Richard Wade-Martins [DESCRIPTION] This protocol, carried out according to manufacturer’s instructions uses a classical ELISA experimental design with sensitive electrochemical detection to provide...
["[Harvesting media and cell lysates] Harvest conditioned media from cells grown in a full-area 96 well plate, being careful not to disturb the cell monolayer. Add 30 uL RIPA lysis buffer (0.5% Triton X-100, 10 mM Tris/HCl, pH 8.0, 1 mM EDTA, 0.5 mM EDTA,0.1% sodium dodecyl sulfate, 0.02% sodium deoxycholate, and (140 ...
44,584
Drug Sensitivity Assays of Human Cancer Organoid Cultures
4
dx.doi.org/10.17504/protocols.io.bpsgmnbw
https://www.protocols.io/view/drug-sensitivity-assays-of-human-cancer-organoid-c-bpsgmnbw
Hayley E. Francies, Andrew Barthorpe, Anne McLaren-Douglas, William J. Barendt, Mathew J. Garnett
TITLE: Drug Sensitivity Assays of Human Cancer Organoid Cultures AUTHORS: Hayley E. Francies, Andrew Barthorpe, Anne McLaren-Douglas, William J. Barendt, Mathew J. Garnett [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Drug sensitivity testing utilizing preclinical disease models such as cancer cel...
["[3.1 Preparation of Drug Stocks and Drug Plates]\nDesign the layout of your drug plate (see Note 4 and Fig. 2).", "[3.1 Preparation of Drug Stocks and Drug Plates]\nDrugs are reconstituted in and stored frozen at or ideally in StoragePods® (Roylan Developments) kept at , providing a moisture-free, low-oxygen enviro...
43,649
DNA Extraction for Beetle DNA with Qiagen DNeasy
3
dx.doi.org/10.17504/protocols.io.bnu9mez6
https://www.protocols.io/view/dna-extraction-for-beetle-dna-with-qiagen-dneasy-bnu9mez6
Demian F Gomez, Caroline Storer
TITLE: DNA Extraction for Beetle DNA with Qiagen DNeasy AUTHORS: Demian F Gomez, Caroline Storer [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The purpose of this protocol is to extract DNA from beetles using the DNeasy protocol with additional notes to the manufacturer guidelines.</div><div class...
[]
28,864
Frequently sampled Insulin glucose tolerance test
null
dx.doi.org/10.17504/protocols.io.8e8hthw
null
Timothy Nichols, David Clemmons
TITLE: Frequently sampled Insulin glucose tolerance test AUTHORS: Timothy Nichols, David Clemmons [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This assay is used by the DiaComp to measure glucose tolerance and insul...
["FSIGT or Bergman analysis Pigs are studied after an overnight fast. The food intake of the animals is monitored for 3 days prior to the fast to ensure adequate carbohydrate intake. Two intravenous catheters are placed, one for sampling and one for infusing glucose and insulin. A bolus of glucose (0.3 gm/kg) is admini...
78,725
3D Mesh Cleanup Tutorial: Fossil Plant Cupule (Beginner)
1
dx.doi.org/10.17504/protocols.io.j8nlkwyywl5r/v1
https://www.protocols.io/view/3d-mesh-cleanup-tutorial-fossil-plant-cupule-begin-cq5dvy26
Elizabeth G. Clark, Kelsey M Jenkins, Craig R Brodersen
TITLE: 3D Mesh Cleanup Tutorial: Fossil Plant Cupule (Beginner) AUTHORS: Elizabeth G. Clark, Kelsey M Jenkins, Craig R Brodersen [DESCRIPTION] This protocol details 3D mesh cleanup tutorial of fossil plant cupule (Beginner). [GUIDELINES] Skills developed: Basic mesh editing 3D meshes of fossil specimens extracted d...
["[Part 1: The Meshlab Interface] Open Meshlab and import “Cupule before.obj” into Meshlab via File > Import Mesh. Alternatively, drag the file into the project window (the large purple box with the crosshairs) if using Mac.", "[Part 1: The Meshlab Interface] The mesh now appears in the project window. You can rotate i...
null
null
null
dx.doi.org/10.17504/protocols.io.fjrbkm6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
83,660
Baiting Pythium myriotylum from Infested Soil
4
null
https://www.protocols.click/view/baiting-pythium-myriotylum-from-infested-soil-cvxkw7kw
Nimalka M Weerasuriya
TITLE: Baiting Pythium myriotylum from Infested Soil AUTHORS: Nimalka M Weerasuriya [DESCRIPTION] Baiting Pythium from Ft. Cobb or other infested soil. [STEPS] SECTION: Preparation 1. Prep P5ARP Plates 1-2 days before plating. Prepare working culture plates (CMA, PDA + amp) up to 1 week before plating. SECTION: Tr...
["[Preparation] Prep P5ARP Plates 1-2 days before plating.\nPrepare working culture plates (CMA, PDA + amp) up to 1 week before plating.", "[Transfer Culture] Check plates after 24-48 h.", "[Oospore Check] Check for \"gold coin\" oospores at plate edges to indicate Pythium myriotylum. \nUse CMA for oospore production\n...
15,223
Nodule cleaning
null
dx.doi.org/10.17504/protocols.io.s4xegxn
null
null
TITLE: Nodule cleaning AUTHORS: [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Adapted for high throughput of small samples for sequencing.</div><div class = "text-block">Based on doi:10.1371/journal.pone.0047677</div></div> [STEPS] ?. Cleanly cut nodules from plant roots and drop into a 2ml Eppe...
["Cleanly cut nodules from plant roots and drop into a 2ml Eppendorf tube per plant.", "Add 1ml 100% EtOH to each tube", "Incubate for 5 seconds", "Remove as much liquid as possible without removing loose nodules", "Add 1ml 6% sodium hypochlorite", "Remove as much liquid as possible", "Add 2ml sterile water", "Remove a...
92,909
Single particle analysis of α-Synuclein fibrils
1
dx.doi.org/10.17504/protocols.io.81wgbxozylpk/v1
https://www.protocols.io/view/single-particle-analysis-of-synuclein-fibrils-c6ymzfu6
Kevin Sicking, Rubén Fernández-Busnadiego
TITLE: Single particle analysis of α-Synuclein fibrils AUTHORS: Kevin Sicking, Rubén Fernández-Busnadiego [DESCRIPTION] Aggregates of α-Synuclein (α-Syn) have been implicated in the pathogenesis of a spectrum of disorders, including Parkinson’s disease, multiple system atrophy, and dementia with Lewy bodies. These agg...
["[Preparation of α-Syn Pre-Formed Fibrils from α-Syn monomers] To produce pre-formed fibrils for structural analysis, adhere to the standardized protocol outlined here: [Protocol for Generation of Pre-formed Fibrils](https://www.protocols.io/view/protocol-forgeneration-of-pre-formed-fibrils-from-rm7vz3ezxgx1/v1).", "[...
62,441
Next Optimal Male Enhancement - (Consumers Warning!) Don’t Buy Until Read!
3
dx.doi.org/10.17504/protocols.io.3byl4bxz8vo5/v1
https://www.protocols.io/view/next-optimal-male-enhancement-consumers-warning-do-b88hrzt6
H Douglas Morris
TITLE: Next Optimal Male Enhancement - (Consumers Warning!) Don’t Buy Until Read! AUTHORS: H Douglas Morris [DESCRIPTION] Plans with high deductibles will only cost you a lot if you actually use medical services. [STEPS]
[]
107,204
Indirect Co-Culture Assay using Boyden Chambers
0
dx.doi.org/10.17504/protocols.io.5jyl82om7l2w/v1
https://www.protocols.io/view/indirect-co-culture-assay-using-boyden-chambers-dkxc4xiw
Bianca Cruz Pachane, Heloisa Sobreiro Selistre de Araujo
TITLE: Indirect Co-Culture Assay using Boyden Chambers AUTHORS: Bianca Cruz Pachane, Heloisa Sobreiro Selistre de Araujo [DESCRIPTION] Here, we describe a transwell assay using Boyden chambers as an indirect co-culture method, where two cell lines are grown together but separated by a porous membrane. This assay was u...
["[Preparation of Gelatin-Coated Coverslips] In preparation: \nClean round glass coverslips (13 mm ø) with 70% ethanol wipes before use. Maintain slips in a clean container.\nPrepare a 0.5 % (v/v)and keep at 4 °C protected from light.", "[Preparation of Gelatin-Coated Coverslips] Drop coverslips atop the droplets and i...
63,359
Polychromatic UV Dose Determination
1
null
https://www.protocols.io/view/polychromatic-uv-dose-determination-b947r8zn
Daniel Ma, NATALIE HULL
TITLE: Polychromatic UV Dose Determination AUTHORS: Daniel Ma, NATALIE HULL [DESCRIPTION] For polychromatic UV light sources, UV irradiance throughout the sample depth can be calculated by changing variables the “Germicidal Fluence (UV Dose) Calculations for a Low Pressure UV Lamp” obtained from Bolton Photoscience...
["[Introduction] The purpose of this protocol is to document the steps used for determination of UV doses for polychromatic UV sources such as UV LEDs and medium pressure mercury lamps. The method is modified from Bolton and Linden (2003). Gather equipment and materials. Sterilize materials and prepare agar for enumera...
86,705
Levodopa-induced dyskinesia mouse model
4
dx.doi.org/10.17504/protocols.io.8epv5xxb5g1b/v1
https://www.protocols.io/view/levodopa-induced-dyskinesia-mouse-model-cywrxxd6
Beatriz E Nielsen
TITLE: Levodopa-induced dyskinesia mouse model AUTHORS: Beatriz E Nielsen [DESCRIPTION] This protocol describes a mouse model of levodopa-induced dyskinesia (LID) and the behavioral assessment of its motor deficits. It includes open field locomotor activity and scoring of abnormal involuntary movements (AIMs). [STEP...
["[Levodopa-induced dyskinesia mouse model development] Generate unilaterally high dose 6-OHDA-lesioned mice (protocol linked below):", "[Levodopa-induced dyskinesia mouse model development] 3 weeks after 6-OHDA injections, verify unilateral dopamine depletion by assessing forelimb use assymetry in the cylinder test (p...
85,814
WU sn-prep Protocol for solid tumors- ATAC v2.7
1
dx.doi.org/10.17504/protocols.io.e6nvwdkx2lmk/v1
https://www.protocols.io/view/wu-sn-prep-protocol-for-solid-tumors-atac-v2-7-cx2wxqfe
Wagma Caravan, Reyka Jayasinghe, Nataly Naser Al Deen
TITLE: WU sn-prep Protocol for solid tumors- ATAC v2.7 AUTHORS: Wagma Caravan, Reyka Jayasinghe, Nataly Naser Al Deen [DESCRIPTION] Single nuclei dissociation protocol for snATAC-seq [STEPS] SECTION: Reagents and Tools 1. 1x Lysis buffer (2mL): 10mM Tris-HCl (pH 7.4) (Thermo; 15567027), 20μL 10mM NaCl (Thermo; AM9759...
["[Reagents and Tools] 1x Lysis buffer (2mL):\n10mM Tris-HCl (pH 7.4) (Thermo; 15567027), 20μL\n10mM NaCl (Thermo; AM9759), 4μL\n3 mM MgCl2 (Thermo; AM9530G), 6μL\nNP-40 substitute (Sigma, 74385-1L), 2μL\n1 M DTT (Sigma, 646563), 2μL\nNuclease Free Water (Invitrogen, AM9937), 1.966mL", "[Reagents and Tools] Lysis Dilut...
null
null
null
dx.doi.org/10.17504/protocols.io.gs4bwgw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol summarizes the process of primer design for assembly-based cloning methods, such as Gibson or AQUA-cloning. </p> <p>Well-designed primers are necessary for efficient cloning of any kind.<strong> </strong></p> <p>This protocol can also be used to design primers f...
[]
48,064
Multiplex Immunofluorescence on Fresh Frozen Tissue-V2
1
dx.doi.org/10.17504/protocols.io.bs68nhhw
https://www.protocols.io/view/multiplex-immunofluorescence-on-fresh-frozen-tissu-bs68nhhw
Maya Brewer, Liz McDonough, Yuantee Zhu, Elizabeth Neumann, Mark De Caestecker, Danielle Gutierrez, Jeff Spraggins
TITLE: Multiplex Immunofluorescence on Fresh Frozen Tissue-V2 AUTHORS: Maya Brewer, Liz McDonough, Yuantee Zhu, Elizabeth Neumann, Mark De Caestecker, Danielle Gutierrez, Jeff Spraggins [DESCRIPTION] Scope: To describe the procedure for multiple cycles of immunofluorescence on human kidney tissue embedded in c...
["[Immunofluorescence] Place frozen slides from the -80˚C freezer directly into the formalin, and post-fix for 5 minutes.", "[Immunofluorescence] Pour used formalin into the appropriate waste container, then fill jar with 1X PBS and wash sections for 5 minutes four times.", "[Immunofluorescence] Using a hydrophobic pen...
null
null
null
dx.doi.org/10.17504/protocols.io.fz4bp8w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Now includes media used for phosphate limitation experiments.</p> [GUIDELINES] <p>All media should be autoclaved unless otherwise noted</p> <p> </p> <p><u>Zobell plates, 100% nutrient (1 L) – for growing host; ~15 mL/plate</u></p> <p>26 g sea salts</p> <p>1 g yeast extract</...
[]
69,611
Passage cells
4
dx.doi.org/10.17504/protocols.io.n92ldpbd7l5b/v1
https://www.protocols.io/view/passage-cells-cf8jtrun
Adita Ayu Permanasari
TITLE: Passage cells AUTHORS: Adita Ayu Permanasari [DESCRIPTION] Passage Cells [STEPS] 1. Observe cells under microscope 2. Wipe outside surface of cell culture dish with ethanol cotton and bring it inside Biosafety Cabinet (BSC) 3. Bring medium, PBS, Trypsin-EDTA 2x to BSC 4. Bring autopipetter, disposable pipet (1...
["Observe cells under microscope", "Wipe outside surface of cell culture dish with ethanol cotton and bring it inside Biosafety Cabinet (BSC)", "Bring medium, PBS, Trypsin-EDTA 2x to BSC", "Bring autopipetter, disposable pipet (10ml, 2ml), one 15ml tube to BSC", "Discard old medium", "Rinse the bottom of dish with 10ml...
53,631
Genomic DNA extraction from the diatom Pseudo-nitzschia multistriata for Illumina sequencing
1
dx.doi.org/10.17504/protocols.io.byk7puzn
https://www.protocols.io/view/genomic-dna-extraction-from-the-diatom-pseudo-nitz-byk7puzn
Francesco Manfellotto, Monia Teresa Russo, Pina Marotta, Rossella Annunziata, Anna Santin, Antonella Ruggiero, Mariella Ferrante
TITLE: Genomic DNA extraction from the diatom Pseudo-nitzschia multistriata for Illumina sequencing AUTHORS: Francesco Manfellotto, Monia Teresa Russo, Pina Marotta, Rossella Annunziata, Anna Santin, Antonella Ruggiero, Mariella Ferrante [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Genomic...
["Detect the presence/absence of bacteria and collect the cultures as described in: dx.doi.org/10.17504/protocols.io.btt5nnq6", "Resuspend cells with 500 μL of TE buffer (10 mM TrisHCl pH 7.6 and 1 mM EDTA pH 8.0)", "Add: 400 mg of 0.2-0.3 mm diameter zirconia/silica beads 500 μL phenol (pH 7.8).", "Mix with vortex 3 t...
24,336
BiomekFXp Robot Minipreps (RoboPreps)
null
dx.doi.org/10.17504/protocols.io.3zqgp5w
null
James Angstman
TITLE: BiomekFXp Robot Minipreps (RoboPreps) AUTHORS: James Angstman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">RoboPreps are great for high-throughput cloning and subcloning, especially if you’re going to be doing Maxipreps afterward anyway. RoboPreps must be done in 96 well format as of now, ...
["[Cell Prep]\nPick colonies into into a 96-well assay block and grow at overnight with shaking.\n[LB cultures]\n37 °C", "[Cell Prep]\nPellet the cells by spinning at for 5-15 minutes .", "[Cell Prep]\nRemove the media by quickly inverting the block over a waste container.\nIf intended for later use, you can freeze...
60,103
Human Knee Cartilage Collection Protocol for Single Cell RNAseq
4
dx.doi.org/10.17504/protocols.io.14egn7996v5d/v1
https://www.protocols.io/view/human-knee-cartilage-collection-protocol-for-singl-b6xfrfjn
molmer , Martin Lotz
TITLE: Human Knee Cartilage Collection Protocol for Single Cell RNAseq AUTHORS: molmer , Martin Lotz [DESCRIPTION] Cartilage from the medial femoral condyle is shaved for Single Cell RNAseq processing. The attached image indicates where the cartilage shavings are collected from. [BEFORE_START] Knee blocks are shipp...
["Prepare harvesting area with sterile drapes, tools and gauze. All harvesting is completed within an aseptic environment.", "Wipe down the knee blocks with 95% ethanol prior to opening the joint capsule.", "Once the joint capsule is opened, the femur and tibia are disarticulated. Wet a sterile gauze with DPBS+1% Anti-...
18,603
Analysis of the time evolution of auditory steady-state responses (ASSR) recorded in rats
null
dx.doi.org/10.17504/protocols.io.wejfbcn
null
Pavel Prado, Eduardo Martínez-Montes, Matías Zañartu
TITLE: Analysis of the time evolution of auditory steady-state responses (ASSR) recorded in rats AUTHORS: Pavel Prado, Eduardo Martínez-Montes, Matías Zañartu [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">Auditory steady-state responses (ASSRs) are ...
["[Preparation]\nAnimals are anesthetized with ketamine (75.0 mg/kg, ip) and diazepam (5.0 mg/kg, ip).", "[Preparation]\nSupplemental doses of anesthesia are administered during the experiment at a level sufficient to maintain the animal in an areflexic state.", "[Preparation]\nAtropine sulfate (0.06 mg/kg; im) are adm...
57,672
Introductory Hydra Activities
1
null
https://www.protocols.io/view/introductory-hydra-activities-b4jgqujw
Callen Hyland
TITLE: Introductory Hydra Activities AUTHORS: Callen Hyland [DESCRIPTION] This is a series of introductory lab activities for BIOL309-03: Research Methods. The purpose is to master vocabulary related to Hydra anatomy, become familiar with compound microscopes and dissecting microscopes, and practice microdissections ...
["[Observation Activities] In this activity you will become familiar with the anatomy of Hydra including areas of the body, cell layers, and nematocysts, while practicing the new vocabulary we learned in lecture.\n\nLearning objectives:\nUse a compound microscope to examine prepared slides\nMaster vocabulary related to...
null
null
null
dx.doi.org/10.17504/protocols.io.es4begw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
78,358
Study Population (Part 4 of &#34;Effects of Online Exercise Intervention on Physical and Mental Conditions in Young Adults with Chronic Neck Pain&#34;)
1
dx.doi.org/10.17504/protocols.io.6qpvr4wxbgmk/v1
https://www.protocols.io/view/study-population-part-4-of-34-effects-of-online-ex-cqrwvv7e
Yiting Lin
TITLE: Study Population (Part 4 of &#34;Effects of Online Exercise Intervention on Physical and Mental Conditions in Young Adults with Chronic Neck Pain&#34;) AUTHORS: Yiting Lin [DESCRIPTION] This is a Part 4 of "Effects of Online Exercise Intervention on Physical and Mental Conditions in Young Adults with Chronic Ne...
["[Inclusion Criteria] Adults between 18 and 50 years of age with neck pain (from occiput to 7thcervical vertebra) for at least 3 months.", "[Inclusion Criteria] A score of >= 4/50 on the NDI.", "[Exclusion Criteria] Adults with a history of previous neck surgery, cervical radiculopathy, acute neck injury or fracture."...
67,422
Alpha Extracts Pure Hemp Oil Reviews Canada: Free Trial and Price of Alpha Extracts CBD Oil
3
dx.doi.org/10.17504/protocols.io.x54v9y68pg3e/v1
https://www.protocols.io/view/alpha-extracts-pure-hemp-oil-reviews-canada-free-t-cd36s8re
pure
TITLE: Alpha Extracts Pure Hemp Oil Reviews Canada: Free Trial and Price of Alpha Extracts CBD Oil AUTHORS: pure [DESCRIPTION] Alpha Extracts Pure Hemp Oil [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.hg3b3yn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div class="bibliography-item-copy-text content col-md-12" data-clipboard-target="copy-target-228943496" data-redirect-target="/items/228943496/copy">Van Etten, J. (n.d.). Formulation of Modified Bold's Basal Medium (MBBM). Retrieved from http://ncv.unl.edu/vanettenlab/</div> <d...
[]
90,702
General genotyping of Myzus persicae using PCR with microsatellite markers and gel electrophoresis
4
dx.doi.org/10.17504/protocols.io.bp2l6x29klqe/v2
https://www.protocols.io/view/general-genotyping-of-myzus-persicae-using-pcr-wit-c4tnywme
Mariska M. Beekman, Marcel Dicke, Bas J. Zwaan, Eveline Verhulst, Bart Pannebakker
TITLE: General genotyping of Myzus persicae using PCR with microsatellite markers and gel electrophoresis AUTHORS: Mariska M. Beekman, Marcel Dicke, Bas J. Zwaan, Eveline Verhulst, Bart Pannebakker [DESCRIPTION] This protocol can be used to 'roughly' genotype green peach aphids, Myzus persicae (Sulzer), based on micr...
["[Materials] Chelex and proteinase K based DNA extraction\nPipettes and pipette tips\nEppendorf tubes \nCentrifuge \nHeat block or water bath \nUltrapure water \nChelex 100 resin (Bio-Rad, Hercules, CA, USA)\nProteinase K (20 mg/mL; Promega, Southampton, UK)\n\nPCR\nPipettes and pipette tips \nPCR machine \nPCR tubes ...
55,463
Scaffold Mapping Protocol - Version 1.0
1
dx.doi.org/10.17504/protocols.io.b2efqbbn
https://www.protocols.io/view/scaffold-mapping-protocol-version-1-0-b2efqbbn
Mabelle Lin, Sonia Sharma
TITLE: Scaffold Mapping Protocol - Version 1.0 AUTHORS: Mabelle Lin, Sonia Sharma [DESCRIPTION] The scope of this protocol covers the steps involved in the mapping of image data to organ scaffolds. [STEPS] 1. Check for the availability of segmentation files for image data. Ensure the image data has already been ...
["Check for the availability of segmentation files for image data.\n\nEnsure the image data has already been segmented with MBF software and the MBF XML file is available. If the MBF XML file has not already been uploaded to Pennsieve, advise the investigators to do so. Check the segmentation file to make sure the data...
98,921
MHV Tissue Titering Protocol
0
null
https://www.protocols.io/view/mhv-tissue-titering-protocol-dcuh2wt6
Siddharth Krishnamurthy
TITLE: MHV Tissue Titering Protocol AUTHORS: Siddharth Krishnamurthy [DESCRIPTION] This protocol is for isolating total nucleic acid from soft tissues in mice, for subsequent analysis of Viral RNA levels [STEPS] SECTION: Day -1: Prepare tissue collection tubes and plate 5. Add 650 µLto each bead containing tube SE...
["[Day -1: Prepare tissue collection tubes and plate] Add 650 µLto each bead containing tube", "[Day -1: Prepare tissue collection tubes and plate] Label a sufficient number of 1.1 mL 12-well cluster tubes, and then place them in a new rack.", "[Day -1: Prepare tissue collection tubes and plate] Add 5-10 2.0 mm Zirconi...
108,368
Meat identification protocol
0
null
https://www.protocols.io/view/meat-identification-protocol-dm3q48mw
openbioscience Adrian
TITLE: Meat identification protocol AUTHORS: openbioscience Adrian [DESCRIPTION] Identifying species by identifying unique DNA sequences is common practice. In this case we will identify the presence or absence of pork DNA in a meat sample. We will use the alkaline method for DNA extraction and LAMP method for DNA am...
["[Meat Preparation] Measure 500 mg pork meat without fat. This is our sample (aliquot.", "Measure 1 ml of the liquid meat and add it to a sterile centrifuge tube labeled \"raw sample\". This is done with a 100- 1000 μl micropipette.", "Prepare 5 ml of NaOH 0.2 mol/L (sodium hydroxide or caustic soda). This will be us...
61,415
Stranded Transcript Count Table Generation from Long Reads
1
dx.doi.org/10.17504/protocols.io.5qpvonn2bl4o/v15
https://www.protocols.io/view/stranded-transcript-count-table-generation-from-lo-b78frrtn
David A Eccles
TITLE: Stranded Transcript Count Table Generation from Long Reads AUTHORS: David A Eccles [DESCRIPTION] This protocol is for comparing different samples at the transcript level, using long reads that are mapped to transcripts. Input(s): demultiplexed and oriented fastq files (see protocol Preparing Reads for Strande...
["[Demultiplex Reads] Demultiplex and orient reads as per the protocol Preparing Reads for Stranded Mapping. It is expected that these demultiplexed reads will be split up in the current directory, and coupled with a 'barcode_counts.txt' file. If that's the case, the following should work:\n \nExample expected output:\...
24,197
Simple subtidal rocky reef environmental parameter station
1
null
https://www.protocols.io/view/simple-subtidal-rocky-reef-environmental-parameter-3vdgn26
Gonzalo Bravo, Gregorio Bigatti, Juan Livore
TITLE: Simple subtidal rocky reef environmental parameter station AUTHORS: Gonzalo Bravo, Gregorio Bigatti, Juan Livore [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">This is a simple system that can be deployed on rocky reefs to measure environmenta...
["[PREPARATION OF SEDIMENT TRAPS]\nPVC tubes of 50 mm diameter were cut into 200 mm long pieces. One of the extremes of the tube was covered by fixing a PVC cup in order to perfectly close that end.Each trap was identified with ID number (Figure 1A). n=3 traps in each reef .130 cm Iron bars (8 mm diameter) were prepare...
48,837
Untargeted Top-down Proteomics by CZE-MS/MS on Eclipse
1
dx.doi.org/10.17504/protocols.io.btxdnpi6
https://www.protocols.io/view/untargeted-top-down-proteomics-by-cze-ms-ms-on-ecl-btxdnpi6
Kevin Jooß, Bryon Drown, Rafael Melani, Kelleher Research Group
TITLE: Untargeted Top-down Proteomics by CZE-MS/MS on Eclipse AUTHORS: Kevin Jooß, Bryon Drown, Rafael Melani, Kelleher Research Group [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Describes the CZE-MS/MS data acquisition procedure for top-down proteomics samples using the Thermo Scientific Orbit...
["Samples were analyzed on a Thermo Scientific Orbitrap Eclipse Tribrid mass spectrometer in line with a Sciex CESI 8000 Plus", "Samples were prepared in ( ) 0.3% acetic acid and transferred to sample vials.\n10 µl", "Prior each sample injection, the capillary is flushed with 0.1 M HCl, filled with new background elect...
40,315
Preparation of a protein-LG conjugated to horseradish peroxidase by the periodate method.
6
dx.doi.org/10.17504/protocols.io.bjk3kkyn
https://www.protocols.io/view/preparation-of-a-protein-lg-conjugated-to-horsera-bjk3kkyn
Angel Justiz-Vaillant
TITLE: Preparation of a protein-LG conjugated to horseradish peroxidase by the periodate method. AUTHORS: Angel Justiz-Vaillant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">SpLG comprises of 4 Ig-binding domains of SpL and 2 IgG Fc-binding SpG domains [1]. This hybrid molecule was found to bind...
["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 (SpL) with an equal amount (500 micrograms) of a mix of horseradish peroxidase-sodium periodate. On the other ...
63,623
2-step PCR mixture and conditions (Barcoded-head primers for seqs pooling)
4
null
https://www.protocols.io/view/2-step-pcr-mixture-and-conditions-barcoded-head-pr-cadfsa3n
Yin-Tse Huang
TITLE: 2-step PCR mixture and conditions (Barcoded-head primers for seqs pooling) AUTHORS: Yin-Tse Huang [DESCRIPTION] PCR mixture and condition (2X SUPERGREEN PCR MASTER MIX) [STEPS] 1. Wear glove, clean up the working bench w. 1% bleach SECTION: For 1&#x27; PCR head-primers 2. Prepare 1' PCR master mixutre for hea...
["Wear glove, clean up the working bench w. 1% bleach", "[For 1&#x27; PCR head-primers] Prepare 1' PCR master mixutre for head-primers (prepare 1.2X of solutions for pipetting error if needed)\n \nPCR mixture for head-primers for each reaction", "[For 1&#x27; PCR head-primers] Mix the 1' PCR master mixture gently by pi...
84,651
Microscopy-based bead assay
4
dx.doi.org/10.17504/protocols.io.14egn38pzl5d/v1
https://www.protocols.io/view/microscopy-based-bead-assay-cwwjxfcn
Elias Adriaenssens
TITLE: Microscopy-based bead assay AUTHORS: Elias Adriaenssens [DESCRIPTION] This protocol describes the microscopy-based bead assay. [STEPS] SECTION: Microscopy-based bead assay 1. We use Glutathione Sepharose 4B beads (GE Healthcare) to bind GST-tagged bait proteins. SECTION: Microscopy-based bead assay 2. To this ...
["[Microscopy-based bead assay] We use Glutathione Sepharose 4B beads (GE Healthcare) to bind GST-tagged bait proteins.", "[Microscopy-based bead assay] To this end, wash 20 µL of beads twice with dH2O and equilibrate with bead assay\nbuffer.", "[Microscopy-based bead assay] Then, resuspend beads in 40 µL bead assay bu...
95,560
Sleep Data Analysis
4
null
https://www.protocols.io/view/sleep-data-analysis-c9jgz4jw
daniel.dautan, Per Svenningsson
TITLE: Sleep Data Analysis AUTHORS: daniel.dautan, Per Svenningsson [DESCRIPTION] Sleep data analysis using Neuroscore V3.0 software. [STEPS] SECTION: Visual inspection and stage definitions 1. Import sleep data files into Neuroscore V3.0 software and visually inspect data. Exclude data with interruptions, noise, or...
["[Visual inspection and stage definitions] Import sleep data files into Neuroscore V3.0 software and visually inspect data.\n\nExclude data with interruptions, noise, or issues related to battery status from further analyses.\n\nUse the DSI-provided \"sleep scoring 2\" script for analysis, incorporating EEG, EMG, and ...
59,233
PBMC isolation
4
dx.doi.org/10.17504/protocols.io.b539q8r6
https://www.protocols.io/view/pbmc-isolation-b539q8r6
Michael Morgan
TITLE: PBMC isolation AUTHORS: Michael Morgan [DESCRIPTION] Isolation of PBMCs from fresh whole human blood in Lithium Heparin blood tubes. [STEPS] SECTION: Lymphocyte separation 1. - Start with everything at room temperature: SECTION: Lymphocyte separation 2. - Add 15 mL Ficoll-Paque Plus to each SepM...
["[Lymphocyte separation] - Start with everything at room temperature:", "[Lymphocyte separation] - Add 15 mL Ficoll-Paque Plus to each SepMate tube by pipetting directly into centre of plastic separator (try not to introduce a lot of air beneath the separator)", "[Lymphocyte separation] - Pool anti-c...
null
null
null
dx.doi.org/10.17504/protocols.io.cutwwm
null
null
TITLE: No Title AUTHORS: [GUIDELINES] The following is a list of antibody concentrations for use with the protocol 'Immunohistochemistry - Drosophila Embryo'<br />Custom antibodies were provided by the published authors.<br /><br />DSHB Antibodies<br /><table style='border-collapse: collapse; width: 853pt;' border='0...
[]
53,701
ADE 2022 Day 1: Background and Fieldwork
3
dx.doi.org/10.17504/protocols.io.bp2l6b67zgqe/v2
https://www.protocols.io/view/ade-2022-day-1-background-and-fieldwork-bypdpvi6
Tom Little
TITLE: ADE 2022 Day 1: Background and Fieldwork AUTHORS: Tom Little [DESCRIPTION] ADE 2022 description of work to be done on Day 1 of the practical work [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.h5eb83e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol describes methods for a mouse xenograft study designed to measure in vivo BiTE® efficacy against antigen-expressing human tumor cells. In a modified version, the protocol is used to measure BiTE®-mediated bystander killing. Mixtures of human EGFR-positive (unlab...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.srred56
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol describes how to deliver ribonucleoprotein (RNP) complexes that consist of purified Cas9 nuclease duplexed with chemically modified synthetic single guide RNA (sgRNA) to induced pluripotent stem (iPS) cells derived from human fibroblasts. RNP delivery is accomplish...
["[Pre-Nucleofection - Seed Cells] Culture iPS cells on Matrigel® Matrix-coated plates until they are semiconfluent.", "[Setup & Nucleofection - Prepare Destination Plate] Coat a new 6-well plate with Matrigel® Matrix and incubate according to the manufacturer’s instructions.", "[Setup & Nucleofection - Prepare Destina...
70,931
Luciferase Activity Assay for Neurospora crassa
3
dx.doi.org/10.17504/protocols.io.q26g7y883gwz/v1
https://www.protocols.io/view/luciferase-activity-assay-for-neurospora-crassa-chhtt36n
kdcastillo
TITLE: Luciferase Activity Assay for Neurospora crassa AUTHORS: kdcastillo [DESCRIPTION] Here, we describe an assay for screening for luciferase activity and performing a circadian luciferase rhythm assay in the filamentous fungus Neurospora crassa. [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.s5meg46
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This plasmid is designed to integrate within the small subunit ribosomal RNA gene region of <em>B. saltans</em>. It carries 1 kb of the <em>B. saltans</em> small subunit ribosomal RNA gene. To construct this cassette, we first sequenced the complete ribosomal operon of <em>B....
[]
52,747
Plasmid construction
4
dx.doi.org/10.17504/protocols.io.bxrjpm4n
https://www.protocols.io/view/plasmid-construction-bxrjpm4n
Chunmei Chang
TITLE: Plasmid construction AUTHORS: Chunmei Chang [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">plasmid construction</div></div> [STEPS] ?. Purify the DNA from gel using a Gel extraction kit (Bio Basic). ?. Linearize the backbone with restriction enzymes (NEB). ?. Amplify the insert gene fragme...
["Purify the DNA from gel using a Gel extraction kit (Bio Basic).", "Linearize the backbone with restriction enzymes (NEB).", "Amplify the insert gene fragment by PCR with primers including 21 nt of overlapping sequence with the target gene.", "Run PCR products and linearized backbone in an agarose gel to confirm the ...
91,038
Preparation of ex-vivo brain slices for physiology experiments
4
dx.doi.org/10.17504/protocols.io.dm6gp328jvzp/v2
https://www.protocols.io/view/preparation-of-ex-vivo-brain-slices-for-physiology-c456yy9e
enrico.zampese
TITLE: Preparation of ex-vivo brain slices for physiology experiments AUTHORS: enrico.zampese [DESCRIPTION] This protocols describes the procedure to obtain ex-vivo mouse brain slices with a vibratome. This protocol is originally meant to collect mibrain coronal slices or coronal/parasagittal striatal slices, but can ...
["[Before the procedure:] A few hours/day before the procedure it is recommended to prepare some slicing solution and freeze it (~200ml).", "[Before the procedure:] Break the pre-frozen slicing solution into a large beaker and add freshly-made slicing solution. With the help of the immersion blender transform the froze...
92,468
Quantitative real-time PCR
4
dx.doi.org/10.17504/protocols.io.e6nvwd4o2lmk/v1
https://www.protocols.io/view/quantitative-real-time-pcr-c6iuzcew
Tatiana Tkatch
TITLE: Quantitative real-time PCR AUTHORS: Tatiana Tkatch [DESCRIPTION] Quantitative real-time PCR protocol used for Day et al. [STEPS] SECTION: gDNA digestion 1. -For each sample prepare 10ul gDNA digestion reaction mix according to SuperScript IV VILO Master Mix protocol (see pdf attached or substeps below). SECT...
["[gDNA digestion] -For each sample prepare 10ul gDNA digestion reaction mix according to SuperScript IV VILO Master Mix protocol (see pdf attached or substeps below).", "[gDNA digestion] Digest gDNA for 2 min min at 37 °C .", "[gDNA digestion] Place the tubes on ice.", "[gDNA digestion] Add SuperScript IV VILO Master ...
null
null
null
dx.doi.org/10.17504/protocols.io.jwacpae
null
null
TITLE: No Title AUTHORS: [BEFORE_START] <p>Prepare primary stock solutions (in dH<sub>2</sub>O) of the following:</p> <ul> <li>NaNO<sub>3</sub>: 75.00 g/L</li> <li>NaH<sub>2</sub>PO<sub>4</sub>·H<sub>2</sub>O: 5.00 g/L</li> <li>Na<sub>2</sub>SiO<sub>3</sub>·9HO: 30.00 g/L (Note: this will only be used if phytoplankto...
[]
88,174
DNA barcoding on Oxford Nanopore: multiplexing up to 24 x 96-well plates
4
dx.doi.org/10.17504/protocols.io.rm7vzx6n4gx1/v1
https://www.protocols.io/view/dna-barcoding-on-oxford-nanopore-multiplexing-up-t-c2cnyave
Robin Floyd, Sean Prosser, Saeideh Jafarpour
TITLE: DNA barcoding on Oxford Nanopore: multiplexing up to 24 x 96-well plates AUTHORS: Robin Floyd, Sean Prosser, Saeideh Jafarpour [DESCRIPTION] This protocol describes laboratory methods for sequencing a standard COI marker (i.e. DNA barcoding), multiplexing up to 2,280 specimens (24 x 96 well plates, with one ne...
["[1. Prepare Indexed PCR Plates] To prepare the partial mastermix (no primers), combine the following components in a sterile bottle of at least 250 mL volume: \n \n\nThis volume is calculated to make a slight excess of partial PCR mix, more than sufficient for 8 reactions x 96 wells x 24 plates.", "[1. Prepare Indexe...
81,094
The derived positive meaning of physical activity for people with a diabetic foot ulcer; a scoping review protocol
1
dx.doi.org/10.17504/protocols.io.x54v9dx4qg3e/v1
https://www.protocols.click/view/the-derived-positive-meaning-of-physical-activity-ctfewjje
Birgit Rasmussen, Lisbeth Uhrenfeldt
TITLE: The derived positive meaning of physical activity for people with a diabetic foot ulcer; a scoping review protocol AUTHORS: Birgit Rasmussen, Lisbeth Uhrenfeldt [DESCRIPTION] Objectives This scoping review aims to explore the derived positive meaning of physical activity in persons with diabetes who have been i...
["[General Information] The derived positive meaning of physical activity for people with a diabetic foot ulcer; a scoping review protocol", "[General Information] Authors: Rasmussen B, Uhrenfeldt L", "[Background] For a person with diabetes mellitus the treatment of a diabetic foot ulcer entails off-loading treatment ...
40,902
ELISA for measurement of serum macrophage migration inhibitory factor (MIF). 
6
dx.doi.org/10.17504/protocols.io.bj7ekrje
https://www.protocols.io/view/elisa-for-measurement-of-serum-macrophage-migrati-bj7ekrje
Angel Justiz-Vaillant, Belkis Ferrer-Cosme
TITLE: ELISA for measurement of serum macrophage migration inhibitory factor (MIF).  AUTHORS: Angel Justiz-Vaillant, Belkis Ferrer-Cosme [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. An anti-human MIF coating antibody is adsorbed onto the microwells by incubation overnight at 4°C with carbonate...
["An anti-human MIF coating antibody is adsorbed onto the microwells by incubation overnight at 4°C with carbonate-bicarbonate buffer.", "Add 50 µl of human serum or plasma. Human MIF present in the serum or plasma binds to antibodies adsorbed into the microwells.", "The microplate is blocked with 3% non-fat milk-PBS b...
null
null
null
dx.doi.org/10.17504/protocols.io.fp3bmqn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This is a collection of Non-Interfering™ (NI) Protein Assay protocols from G-Biosciences.</p> [STEPS] ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.prrdm56
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>A-2. <u>ISCO Sampling Protocol</u></strong></p> <p>River and estuary samples will be collected using an automated ISCO sampler using the following protocol:</p> <p> </p> <ul> <li>Sampler Description:</li> </ul> <ul> <li>A Teledyne ISCO 3700 full size portable sequenti...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.gzsbx6e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol to prep plasmid DNA from yeast cells. Preps both plasmid and genomic DNA. Protocol for prepping an entire plate; for selection of correctly assembled plasmids, the prepped DNA is transformed into <em>E. coli</em>, and transformation can be followed by colony PCR to s...
[]
28,895
Qualitative paper ELONA test
null
dx.doi.org/10.17504/protocols.io.8f7htrn
null
Manuela de las Casas
TITLE: Qualitative paper ELONA test AUTHORS: Manuela de las Casas [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The aim of this protocol is to test the best method that identifies the efficiency of the detection system that will be present in the strips. 3 methods are tested and compared.</div><di...
["[Preparing the nitrocellulose strip]\nSet the hot plate to 100ºC or at least warm enough to melt the wax. Cut a small amount from one of the wax pencils and place it on a Petri dish. Set the Petri dish on the hot plate and wait for the wax to melt. Cut a strip from the nitrocellulose sheet with the desired size.Once...
null
null
null
dx.doi.org/10.17504/protocols.io.qyndxve
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>e</p> <p>qe</p> <p> </p> <p>wqeewq</p> <p> </p> <p>we</p> <p>wq</p> <p> </p> <p> </p> <p>e</p> <p>wq</p> <p>ew</p> <p> </p> <p>wqwq</p> <p> </p> <p>wqe</p> <p>wqeqeqe</p> <p>wq</p> <p>e</p> <p>wqe</p> <p>wqe</p> <p>q</p> <p>ewqe</p> <p>wqe</p> [BEFORE_START] <p>qwd</p> <p>qd...
[]
46,887
Mass Spectrometry analysis and Molecular MS/MS network
4
dx.doi.org/10.17504/protocols.io.br2fm8bn
https://www.protocols.io/view/mass-spectrometry-analysis-and-molecular-ms-ms-net-br2fm8bn
Rene Flores Clavo, Cristian Daniel Asmat Ortega, Nataly Ruiz Quinones
TITLE: Mass Spectrometry analysis and Molecular MS/MS network AUTHORS: Rene Flores Clavo, Cristian Daniel Asmat Ortega, Nataly Ruiz Quinones [STEPS] ?. [UHPLC ] Resuspend the dry extract in 1 mL of methanol (HPLC grade) and dilute 100 µL in 900 µL of methanol (HPLC grade). ?. [UHPLC ] Filter the final solution using a...
["[UHPLC ]\nResuspend the dry extract in 1 mL of methanol (HPLC grade) and dilute 100 µL in 900 µL of methanol (HPLC grade).", "[UHPLC ]\nFilter the final solution using a syringe-filter into vials", "[UHPLC ]\nPerform Ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS) analyses in a Thermo Scientifi...
58,620
Phytophthora pluvialis culture cultivation, maintenance, and detached needle assay protocol
4
dx.doi.org/10.17504/protocols.io.kqdg3p9d7l25/v1
https://www.protocols.io/view/phytophthora-pluvialis-culture-cultivation-mainten-b5g4q3yw
Sophie Eccersall, Leann Seesom Vinson, Rebecca McDougal, Claudia-Nicole Meisrimler
TITLE: Phytophthora pluvialis culture cultivation, maintenance, and detached needle assay protocol AUTHORS: Sophie Eccersall, Leann Seesom Vinson, Rebecca McDougal, Claudia-Nicole Meisrimler [DESCRIPTION] Phytophthora pluvialis is an oomycete that primarily infects Pinus radiata and Pseudotsuga menziesii causing the d...
["[A. Growth of P. pluvialis from existing culture plates] Grow P. pluvialis isolate on 20% cV8 agar (modified from Jeffers, 2006) at ~18 ˚C.", "[A. Growth of P. pluvialis from existing culture plates] Seal plates with parafilm, and store upside down in an incubator at 17-21 ˚C in the dark. For zoospore production, all...
79,363
Aqueous (SBiP) Delipidation of a Whole Mouse Brain
1
dx.doi.org/10.17504/protocols.io.n2bvj81mwgk5/v1
https://www.protocols.click/view/aqueous-sbip-delipidation-of-a-whole-mouse-brain-crrbv52n
Andrew Recknagel, Kevin Cao, Naveen Ouellette, Molly Logsdon, Judith Baka, Jayaram Chandrashekar
TITLE: Aqueous (SBiP) Delipidation of a Whole Mouse Brain AUTHORS: Andrew Recknagel, Kevin Cao, Naveen Ouellette, Molly Logsdon, Judith Baka, Jayaram Chandrashekar [DESCRIPTION] Aqueous strategies for whole-brain delipidation involve lipid removal via phase separation and detergent washes. SBiP is an aqueous biphasic ...
["[SBiP Delipidation] Use a 20 mL vial for processing an adult mouse brain. All steps in this section are carried out on a carousel rotator . \nReplace solution in vial with 20 mL SBiP for each of the following steps:\nSBiP for 180 min \nSBiP for 360 min \nSBiP \nSBiP for 1440 min \nSBiP for 1440 min\nSBiP for 1440 ...
106,989
Optimized Stereotactic Injection Protocol for Targeting the Locus Coeruleus with Minimal Neurotoxicity
0
dx.doi.org/10.17504/protocols.io.3byl4988ogo5/v1
https://www.protocols.io/view/optimized-stereotactic-injection-protocol-for-targ-dkqm4vu6
Csilla Novák, Rukhshona Kayomova, Matthias Prigge
TITLE: Optimized Stereotactic Injection Protocol for Targeting the Locus Coeruleus with Minimal Neurotoxicity AUTHORS: Csilla Novák, Rukhshona Kayomova, Matthias Prigge [DESCRIPTION] The Locus Coeruleus is a critical brain region known for its vulnerability to mechanical perturbations, neuroinflammation, and axonal di...
["[Animal Preparation] Weigh the mouse to calculate the precise dosage of anesthetic required. Administer ketamine at a dosage of 10 mg/kg and xylazine at a dosage of 20 mg/kg. Alternatively, use isoflurane as approved by your institutional animal care and use protocols. Ensure adherence to all relevant guidelines and ...
38,703
Isolation, Culture, and Maintenance of Patient-Derived Tumor Biopsy
4
null
https://www.protocols.io/view/isolation-culture-and-maintenance-of-patient-deriv-bh2pj8dn
Xiling Shen, Marcos Negrete, Kun Xiang
TITLE: Isolation, Culture, and Maintenance of Patient-Derived Tumor Biopsy AUTHORS: Xiling Shen, Marcos Negrete, Kun Xiang [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is split into 3 sections: collecting tumor cells, passaging, and cryopreserving organoids. </div><div class = "text...
["[Disassociation of Tumor Cells]\nStore tissue samples in cold transport media (). Keep samples on ice at all time and process within\n10 mL", "[Disassociation of Tumor Cells]\nTransfer the tumor biopsy sample (1-2 cm3) and transport media into a petri dish and remove remnant non-tumor tissue with sterile tweezers.\nT...
69,880
Immunochemistry on paraffin sections
4
dx.doi.org/10.17504/protocols.io.kxygx9my4g8j/v1
https://www.protocols.io/view/immunochemistry-on-paraffin-sections-cggyttxw
miquel.vila
TITLE: Immunochemistry on paraffin sections AUTHORS: miquel.vila [DESCRIPTION] Immunochemistry protocol on paraffin-embedded rat brain sections [STEPS] SECTION: 1. Deparaffinization and hydratation : 1. Put the slides 30 min in the incubator at 60ºC. SECTION: 1. Deparaffinization and hydratation : 2. Wash 3x3 min in...
["[1. Deparaffinization and hydratation :] Put the slides 30 min in the incubator at 60ºC.", "[1. Deparaffinization and hydratation :] Wash 3x3 min in Xilen.", "[1. Deparaffinization and hydratation :] Wash 1x10 min in ethanol 100%.", "[1. Deparaffinization and hydratation :] Wash 1x10 min in ethanol 95%.", "[1. Depara...
81,822
NCEM Drop - Paramyxo confirmation, generation of RNP (TM-014)
4
dx.doi.org/10.17504/protocols.io.x54v9dq8pg3e/v1
https://www.protocols.io/view/ncem-drop-paramyxo-confirmation-generation-of-rnp-ct56wq9e
sandra.crameri
TITLE: NCEM Drop - Paramyxo confirmation, generation of RNP (TM-014) AUTHORS: sandra.crameri [DESCRIPTION] Generate RNP to confirm Paramyxoviurs [STEPS] SECTION: HEADER 1. SAN: SPEC No: OPERATOR: SECTION: Conventional 4. Adsorb 10 µL sample to grid 10 min , inspect to ensure sample does not dry out. SECTI...
["[HEADER] SAN:\n\n\n\n\nSPEC No:\n\n\n\n\nOPERATOR:", "[Conventional] Adsorb 10 µL sample to grid 10 min , inspect to ensure sample does not dry out.", "[Conventional] Drain excess sample from grid using filter paper, leave wet.", "[Conventional] Stain 1 min", "Drain & dry using filter paper", "[Generation of RNP] Pr...
63,838
To test fork notification
1
dx.doi.org/10.17504/protocols.io.n92ldzjxxv5b/v2
https://www.protocols.io/view/to-test-fork-notification-caj6scre
Gabriel Gasque
TITLE: To test fork notification AUTHORS: Gabriel Gasque [DESCRIPTION] Test for Fork notifications [STEPS] 1. Step 1 2. Step 2 3. Step 3 4. Step 4
["Step 1", "Step 2", "Step 3", "Step 4"]
101,746
The Impact of World Pandemics on Health Services Management: The Covid 19 Experience in Nigeria and Australia
0
dx.doi.org/10.17504/protocols.io.x54v92ro1l3e/v1
https://www.protocols.io/view/the-impact-of-world-pandemics-on-health-services-m-dfks3kwe
Charles C Okonkwo, Rasheda Khanam, Gavin Beccaria, Ezekiel Uba Nwose
TITLE: The Impact of World Pandemics on Health Services Management: The Covid 19 Experience in Nigeria and Australia AUTHORS: Charles C Okonkwo, Rasheda Khanam, Gavin Beccaria, Ezekiel Uba Nwose [DESCRIPTION] This research aims to evaluate the impact of COVID-19 pandemic on health services management using the COVID-1...
["[Abstract] This research aims to evaluate the impact of COVID-19 pandemic on health services management using the COVID-19 experiences as the major means of measure. Secondary data would be used to compare the Low and Medium-Income Countries (LMIC) and the High-Income Countries (HIC), focusing on health systems in Ni...
33,631
Protocols for the draft genome assembly of the eastern banjo frog Limnodynastes dumerilii dumerilii
2
dx.doi.org/10.17504/protocols.io.bc37iyrn
https://www.protocols.io/view/protocols-for-the-draft-genome-assembly-of-the-eas-bc37iyrn
Qiye Li, Qunfei Guo, Yang Zhou, Huishuang Tan, Terry Bertozzi, Yuanzhen Zhu, Ji Li, Stephen Donnellan, Guojie Zhang
TITLE: Protocols for the draft genome assembly of the eastern banjo frog Limnodynastes dumerilii dumerilii AUTHORS: Qiye Li, Qunfei Guo, Yang Zhou, Huishuang Tan, Terry Bertozzi, Yuanzhen Zhu, Ji Li, Stephen Donnellan, Guojie Zhang [DESCRIPTION] Background: Amphibian genomes are usually challenging to assemble due to ...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ud9es96
null
null
TITLE: No Title AUTHORS: [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 this protocol a...
["[PCR] {\"blocks\":[{\"key\":\"9pc18\",\"text\":\"PCR reactions were run in triplicate 25-\\u03bcl reactions for each sample using 12-basepair Golay barcoded reverse primers (Amaral-Zettler et al., 2009).\\u00a0\\n\\nAmaral-Zettler LA, McCliment EA, Ducklow HW, Huse SM (2009) A Method for Studying Protistan Diversity ...
36,569
Perinatal 2016 Rio Grande - Urinary Incontinence
3
dx.doi.org/10.17504/protocols.io.bfxzjpp6
https://www.protocols.io/view/perinatal-2016-rio-grande-urinary-incontinence-bfxzjpp6
Yuan Hsu, Juraci A Cesar
TITLE: Perinatal 2016 Rio Grande - Urinary Incontinence AUTHORS: Yuan Hsu, Juraci A Cesar [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.pmvdk66
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
85,115
“Skeeter Pheeder”: inexpensive 3D printed, battery-powered blood feeder for mosquitoes and other haematophagous arthropods
1
null
https://www.protocols.click/view/skeeter-pheeder-inexpensive-3d-printed-battery-po-cxc3xiyn
Philip K Stoddard
TITLE: “Skeeter Pheeder”: inexpensive 3D printed, battery-powered blood feeder for mosquitoes and other haematophagous arthropods AUTHORS: Philip K Stoddard [DESCRIPTION] A portable heated feeder for blood-feeding arthropods, the Skeeter Pheeder, has been designed to be constructed in the laboratory using conventional...
["[Materials Needed] See Materials section \nA table is include with details, approximate costs, and possible suppliers.\n\nNote: One can purchase blunt #20 hypodermic needles at a premium price or make them by grinding the tips off sharp hypodermic needles (I used a Dremel hand grinder). The needle shaft should be ab...
86,610
U54 SCENT Pediatric Colonoscopy Tissue Collection Procedure
1
dx.doi.org/10.17504/protocols.io.yxmvm3oznl3p/v2
https://www.protocols.io/view/u54-scent-pediatric-colonoscopy-tissue-collection-cytsxwne
Mary Jordan*
TITLE: U54 SCENT Pediatric Colonoscopy Tissue Collection Procedure AUTHORS: Mary Jordan* [DESCRIPTION] This document outlines the required criteria and acquisition for pediatric colonoscopy colon specimens collected at Duke University Hospital through the Biorepository and Precision Pathology Center (BRPC) in the Depa...
["[Inclusion and Exclusion Criteria] Inclusion Criteria:\n • Healthy patients age 5-18 years of age presenting for outpatient colonoscopy. \n • Screening may wish to focus on the following indications: abdominal pain, constipation, diarrhea, \n bloating/gas, patients with normal labs and/or imaging....
null
null
null
dx.doi.org/10.17504/protocols.io.imucc6w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>March, 2012; based on sample preparation guide v2, catalog # RS-930-1021, part # 15026495 Rev. A, August 2011</p> <p> </p> <p>Modifications: samples in PCR tubes (because a plate/seal will not be used, some numbered steps are skipped); include two rounds of poly-A selection (...
[]
58,840
Nested VP1 PCR and Nanopore Sequencing from Stool and ES Samples v1.11
1
dx.doi.org/10.17504/protocols.io.b5pyq5pw
https://www.protocols.io/view/nested-vp1-pcr-and-nanopore-sequencing-from-stool-b5pyq5pw
Alex Shaw, Manasi Majumdar, Catherine Troman, Joyce Akello, Javier Martin, Nick Grassly
TITLE: Nested VP1 PCR and Nanopore Sequencing from Stool and ES Samples v1.11 AUTHORS: Alex Shaw, Manasi Majumdar, Catherine Troman, Joyce Akello, Javier Martin, Nick Grassly [DESCRIPTION] This protocol is updated from the protocol described in the paper "Rapid and sensitive direct detection and identification o...
["[Nested PCR First Round (PanEV)] Nested PCR First Round (panEV primers):\nPrepare a Master mix using reaction volumes as detailed below, excluding forward primer and the RNA:\n\nForward Primer (5'NTR): [TGGCGGAACCGACTACTTTGGGTG] (Arita et al. 2015)\nReverse Primer (Cre): [TCAATACGGTGTTTGCTCTTGAACTG] (Arita et al....
64,025
Pooling
4
null
https://www.protocols.io/view/pooling-carzsd76
Allyson Hirsch, George Testo
TITLE: Pooling AUTHORS: Allyson Hirsch, George Testo [DESCRIPTION] The throughput of modern sequencers grows higher and higher, allowing hundreds of millions of reads in a single microfluidic chamber. For many purposes this is actually far more than a single library requires. To keep sequencing cost-effective, resear...
["[Preparations] DNA Away and Ethanol the workstation.", "[Preparations] Remove PCR plate(s) from the-20 °C freezer and thaw plates to be pooled while finishing workstation prep.", "[Preparations] Print plate table with volume of each amplicon to be pooled together (equal molar).", "[Preparations] Spin thawed plates do...
55,691
High Throughput SARS-COV-2, PMMOV, and BCoV quantification in settled solids using digital RT-PCR
1
dx.doi.org/10.17504/protocols.io.b2mjqc4n
https://www.protocols.io/view/high-throughput-sars-cov-2-pmmov-and-bcov-quantifi-b2mjqc4n
Aaron Topol (Verily Life Sciences), marlene.wolfe , Brad White (Verily Life Sciences), Krista Wigginton, Alexandria B B Boehm
TITLE: High Throughput SARS-COV-2, PMMOV, and BCoV quantification in settled solids using digital RT-PCR AUTHORS: Aaron Topol (Verily Life Sciences), marlene.wolfe , Brad White (Verily Life Sciences), Krista Wigginton, Alexandria B B Boehm [DESCRIPTION] This process instruction describes the steps for quantitative a...
["[Preparation (both assays)] Retrieve all kit components from the One-Step RT-ddPCR advanced kit for probes from the -20 °C freezer and thaw the components on ice.", "[Preparation (both assays)] Retrieve ddPCR positive control aliquots (50 copies per uL gRNA and 100 copies per µL BCoV and PMMoV gene blocks) from the -...
105,358
Fixation and Immunostaining
0
dx.doi.org/10.17504/protocols.io.rm7vzj2d2lx1/v2
https://www.protocols.io/view/fixation-and-immunostaining-di5n4g5e
Gist Croft, Marta Skowronska
TITLE: Fixation and Immunostaining AUTHORS: Gist Croft, Marta Skowronska [DESCRIPTION] This protocol is based on standard methods for formaldehyde fixation and immunostaining for fluorescence microscopy. However, it contains tested working reagents, advice, and tips. [GUIDELINES] Read all reagent SDS and follow all s...
["[BUFFER PREPARATION] 2 x Fixation Buffer (8% Paraformaldehyde in 2x PBS): Use a 32% paraformaldehyde (PFA) in sealed EMS ampule + 30 mL PBS (+/+), pH 7.4 + 8 mL 10x PBS + 22 mL ddH2O\nOBS: You can use either PBS (+/+) (with Ca+2 and Mg+2) or (-/-). Some might prefer HBSS instead.", "[BUFFER PREPARATION] 1 x Fixation ...
99,670
Podocoryna ACME cell dissociations - draft v1.0, May 13 2024
0
null
https://www.protocols.io/view/podocoryna-acme-cell-dissociations-draft-v1-0-may-ddjw24pe
Michael Connelly
TITLE: Podocoryna ACME cell dissociations - draft v1.0, May 13 2024 AUTHORS: Michael Connelly [DESCRIPTION] This protocol is intended to dissociate and fix cells of the hydrozoan Podocoryna carnea for single-cell RNA sequencing on the 10X Chromium platform following a FACS sorting step. ACME-fixed Podocoryna cells mai...
["[Polyp ACME dissociation] Prepare for polyp dissection by placing a microscope slide with a Podocoryna colony in a glass dish with ~200 mL 0.2 um filtered seawater. Add menthol crystals at a concentration of ~1 g/L to relax the polyps for ~10 minutes.", "[Medusa ACME dissociation] Collect as many medusae as possible ...
86,829
High-Throughput Generation of Single Cell Libraries using SmartSeq2 Plate Assay
1
dx.doi.org/10.17504/protocols.io.8epv5xx54g1b/v1
https://www.protocols.io/view/high-throughput-generation-of-single-cell-librarie-cy2mxyc6
Michael Borja, Tabula Muris Consortium, Shayan Hosseinzadeh
TITLE: High-Throughput Generation of Single Cell Libraries using SmartSeq2 Plate Assay AUTHORS: Michael Borja, Tabula Muris Consortium, Shayan Hosseinzadeh [DESCRIPTION] As part of the larger efforts of Quantitative Cell Science at the Chan Zuckerberg Biohub, single cell plate-based protocols such as the SmartSeq 2 as...
["[Lysis Plate Preparation] Lysis plates were created by dispensing 0.4 μl lysis buffer master mix into 384-well hard-shell PCR plates (Bio- Rad HSP3901) using the SPT LabTech DragonFly. 96-well lysis plates were also prepared with 4 μl lysis buffer. All plates were sealed with Microseal Seal F foil seal and spun down ...
81,996
Protocol of Spotted Fever Rickettsia IgG
4
dx.doi.org/10.17504/protocols.io.bp2l69jdrlqe/v1
https://www.protocols.io/view/protocol-of-spotted-fever-rickettsia-igg-cubkwskw
narankhajid
TITLE: Protocol of Spotted Fever Rickettsia IgG AUTHORS: narankhajid [DESCRIPTION] All serum samples were tested for Rickettsia IgG using a commercial ELISA kit (NovaTec Immunodiagnostica GmbH, Dietzenbach, Germany) according to the manufacturer’s instructions(S9). [STEPS] SECTION: Protocol of Spotted Fever Rickettsi...
["[Protocol of Spotted Fever Rickettsia IgG]", "[Protocol of Spotted Fever Rickettsia IgG] Dispense 100µl controls and diluted samples into their respective wells. Leave well A1 for substrate blank.", "[Protocol of Spotted Fever Rickettsia IgG] Cover wells with the foil supplied in the kit.", "[Protocol of Spotted Feve...
39,565
OptoPlate Calibration protocol
1
dx.doi.org/10.17504/protocols.io.bivmke46
https://www.protocols.io/view/optoplate-calibration-protocol-bivmke46
Edvard Grødem, Kieran Sweeney, Megan N. McClean
TITLE: OptoPlate Calibration protocol AUTHORS: Edvard Grødem, Kieran Sweeney, Megan N. McClean [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Optogenetic systems use light to precisely control and investigate cellular processes. Until recently, there had been few instruments available for applying ...
["[Configuring the optoPlate hardware]\nSection overviewWe assemble the optoPlate as published by Bugaj and Lim, but with two changes to suit the needs of our lab and allow calibration. First, we adapt the optoPlate to accommodate our 96 well optical-bottom plates (Nunc, #265300) via 3D printed adaptors, which can be f...
71,406
Human Liver Tissue Storage Methods For Multiomic Applications
4
dx.doi.org/10.17504/protocols.io.261ge34qdl47/v1
https://www.protocols.io/view/human-liver-tissue-storage-methods-for-multiomic-a-chynt7ve
Diana Nakib, Catia Perciani, Sai Chung, Xue-Zhong Ma, Justin Manuel, Ian McGilvray, Sonya Macparland
TITLE: Human Liver Tissue Storage Methods For Multiomic Applications AUTHORS: Diana Nakib, Catia Perciani, Sai Chung, Xue-Zhong Ma, Justin Manuel, Ian McGilvray, Sonya Macparland [DESCRIPTION] Sampling different areas of the diseased or healthy liver and storing the tissue in a variety of mediums to allow for an in-de...
["[Sample Collection] Collect explanted tissue section in 30mL of HBSS 1x with Ca2+ and Mg2+ in a 50mL conical tube and keep on ice, ideally to process within 30 minutes of removal from patient or removal from the flushed donor organ in preservation solution.", "[Optimal Cutting Temperature (OCT)-Embedding Tissue Secti...
68,850
Modified Arabidopsis Root smRNA FISH Protocol
4
dx.doi.org/10.17504/protocols.io.rm7vzyworlx1/v1
https://www.protocols.io/view/modified-arabidopsis-root-smrna-fish-protocol-cfgstjwe
Susan Duncan, Hans Johansson
TITLE: Modified Arabidopsis Root smRNA FISH Protocol AUTHORS: Susan Duncan, Hans Johansson [DESCRIPTION] Single molecule RNA FISH (smRNA FISH) is an imaging method that labels individual mRNA molecules in cells to facilitate localization and quantitative studies. Here we present a modified protocol for mRNA labelling ...
["[Plant Growth] Sterilize then sow a row of Col-0 Arabidopsis seeds onto half strength Murashige and Skoog Medium (1/2 MS) near the top of a 10 cm square petri plate.", "[Plant Growth] Stratify the seeds at 4 °Cfor two days.", "[Plant Growth] Take the plate out of the cold and place it vertically in a growth cabinet s...
34,783
3D Printing Of Personal Protective Equipment And Repurposing Of Common Household Air Filters
null
dx.doi.org/10.17504/protocols.io.bd77i9rn
null
Giana Schena, Emma Murray
TITLE: 3D Printing Of Personal Protective Equipment And Repurposing Of Common Household Air Filters AUTHORS: Giana Schena, Emma Murray [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>During the height of the COVID-19 outbreak, personal protective ​</span><a href="Https://time.com/5785223/medic...
["[Two Piece Face Mask Printing and Assembly]\nOpen the​ facemask_filter_guard.stl f​ile (currently on scale with the face unit, would also need to be scaled up). a. Support is not necessary b. Guard is in shape of a trapezoid: the face of the guard is 2.5 inches tall with 2.75 inch base length and 1.6 ...
64,178
Wedge sampling of pars petrosa (os temporale) for ancient DNA extraction
1
dx.doi.org/10.17504/protocols.io.261gen8wog47/v1
https://www.protocols.io/view/wedge-sampling-of-pars-petrosa-os-temporale-for-an-cawssfee
Raphaela Stahl, Lena Semerau, Eleftheria Orfanou, Marie Himmel, Franziska Aron, Wolfgang Haak
TITLE: Wedge sampling of pars petrosa (os temporale) for ancient DNA extraction AUTHORS: Raphaela Stahl, Lena Semerau, Eleftheria Orfanou, Marie Himmel, Franziska Aron, Wolfgang Haak [DESCRIPTION] This protocol describes how to obtain bone powder from the pars petrosa of disarticulated ossis temporalis, specifically f...
["[Workstation preparation] Place a sheet of aluminium foil under the hood.\n\nPlace weighing paper, drill bits, and a pair of tooth pliers or forceps on a second sheet of aluminium foil on an easily accessible clean surface outside of the hood. Alternatively, you may place them within the hood but be sure to cover the...
69,549
Integration of malaria and schistosomiasis prevention and control programs: protocol for scoping review
1
dx.doi.org/10.17504/protocols.io.j8nlkwb1xl5r/v1
https://www.protocols.io/view/integration-of-malaria-and-schistosomiasis-prevent-cf6mtrc6
Claudia Duguay, Sydney Raduy, Natacha Protopopoff, Cindy Feng, Alison Krentel, Manisha Kulkarni
TITLE: Integration of malaria and schistosomiasis prevention and control programs: protocol for scoping review AUTHORS: Claudia Duguay, Sydney Raduy, Natacha Protopopoff, Cindy Feng, Alison Krentel, Manisha Kulkarni [DESCRIPTION] Context: It is imperative to build resilient, and sustainable programs to maintain effo...
["[Title and author identification] Integration of malaria and schistosomiasis prevention and control programs: protocol for scoping review\n\nClaudia Duguay1, Sydney Raduy1, Natacha Protopopoff2, Cindy Feng3, Alison Krentel1,4, Manisha A. Kulkarni1\n\n1 School of Epidemiology and Public Health, University of Ottawa, O...