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57,845
RNA purification and cDNA synthesis
1
dx.doi.org/10.17504/protocols.io.ewov1nnzogr2/v1
https://www.protocols.io/view/rna-purification-and-cdna-synthesis-b4qvqvw6
Vicki Deng, Fredrick leon, David Booth
TITLE: RNA purification and cDNA synthesis AUTHORS: Vicki Deng, Fredrick leon, David Booth [DESCRIPTION] This protocol compiles multiple methods for purifying RNA from an S. rosetta lysate and provides a modified reverse transcriptase protocol to robustly synthesize cDNA from transcripts with higher GC content. [STE...
["[Culture cells for lysate] Grow enough culture for ~50x107 cells. \nfor swimming cultures: seed 40 mLacclimated cells at ~8*10^4 cells/mlthen culture at 27 °C for 1440 min \nfor thecate cultures: seed 100 mL acclimated cells in culture plate then culture at 27 °C for 1440 min \n\nNumber of cells doesn't necessarily m...
93,032
Immunofluorescent Staining of Fixed Mouse Brain Tissue Sections
1
dx.doi.org/10.17504/protocols.io.kxygx38owg8j/v1
https://www.protocols.io/view/immunofluorescent-staining-of-fixed-mouse-brain-ti-c64gzgtw
Katerina Rademacher, Ken Nakamura
TITLE: Immunofluorescent Staining of Fixed Mouse Brain Tissue Sections AUTHORS: Katerina Rademacher, Ken Nakamura [DESCRIPTION] This protocol describes steps for immunofluorescent staining of free floating fixed mouse brain tissue sections. [STEPS] 1. Free floating sections in 12-well plates, all steps performed at r...
["Free floating sections in 12-well plates, all steps performed at room temperature.", "2 x 10min PBS washes.", "2 x 10min 0.2% PBS-Triton (PBS-T) washes.", "Block 1hr in blocking buffer.", "Incubate in primary antibody diluted in blocking buffer overnight.", "3 x 10min 0.2% PBS-T washes.", "Incubate in secondary antib...
62,373
Genxz Keto Gummies : Do NOT Buy Until Reading This!
3
dx.doi.org/10.17504/protocols.io.j8nlkknoxl5r/v1
https://www.protocols.io/view/genxz-keto-gummies-do-not-buy-until-reading-this-b86drza6
H A
TITLE: Genxz Keto Gummies : Do NOT Buy Until Reading This! AUTHORS: H A [DESCRIPTION] whilst the majority of your studies will be at one college, the rest of your time will be spent at another university. [STEPS]
[]
106,403
Efficient and precise targeting of the AAVS1 safe harbour locus in hPSCs.
0
null
https://www.protocols.io/view/efficient-and-precise-targeting-of-the-aavs1-safe-dj6b4ran
Dmitry Ovchinnikov
TITLE: Efficient and precise targeting of the AAVS1 safe harbour locus in hPSCs. AUTHORS: Dmitry Ovchinnikov [DESCRIPTION] Stably genetically-modified human pluripotent stem cells (hPSCs) are increasingly being used for studies relying on the consistent expression of the transgene of interest in human stem cells and t...
["[Transfection for gene targeting] 1. Prepare a desired number of wells in a 6-well plate to accommodate the hPSC cell suspension after electroporation, and become \"master\" wells for establishing targeted clones after antibiotic selection. The wells are coated with or similar ECM with 2x higher concentration relati...
57,258
Fast Agarose Gel Electrophoresis-Chem 384/584
1
dx.doi.org/10.17504/protocols.io.b36iqrce
https://www.protocols.io/view/fast-agarose-gel-electrophoresis-chem-384-584-b36iqrce
Ken Christensen, Addgene The Nonprofit Plasmid Repository
TITLE: Fast Agarose Gel Electrophoresis-Chem 384/584 AUTHORS: Ken Christensen, Addgene The Nonprofit Plasmid Repository [DESCRIPTION] This protocol is for agarose gel electrophoresis. To see the full abstract and additional resources, visit the Addgene protocol page. [GUIDELINES] Analyzing your gel Using the DNA ...
["[Pouring a Standard 1% Agarose Gel] Measure .6 g of agarose.", "[Pouring a Standard 1% Agarose Gel] Mix agarose powder with 60 mL 1xTAE in a microwavable flask. See TAE Recipe.", "[Pouring a Standard 1% Agarose Gel] Microwave for 1 min to 3 min until the agarose is completely dissolved (but do not overboil the soluti...
50,920
Thai Casinos For Online Gambling
3
dx.doi.org/10.17504/protocols.io.bvygn7tw
https://www.protocols.io/view/thai-casinos-for-online-gambling-bvygn7tw
Alex george
TITLE: Thai Casinos For Online Gambling AUTHORS: Alex george [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Top casino provide online casino games.</div></div> [STEPS]
[]
84,412
Single cell RNA-seq and ATAC-seq protocol for PBMCs treated with LPS
4
dx.doi.org/10.17504/protocols.io.yxmvm3xznl3p/v1
https://www.protocols.click/view/single-cell-rna-seq-and-atac-seq-protocol-for-pbmc-cwn4xdgw
Francesca Luca
TITLE: Single cell RNA-seq and ATAC-seq protocol for PBMCs treated with LPS AUTHORS: Francesca Luca [DESCRIPTION] Single cell RNA-seq and ATAC-seq protocol for PBMCs treated with LPS [STEPS] 1. Cell Processing – day 1 2. Transfer the selected cryovials from liquid nitrogen into dry ice then store in -80C. The day bef...
["Cell Processing – day 1", "Transfer the selected cryovials from liquid nitrogen into dry ice then store in -80C. The day before seeding the plates (Day 1)", "Day 1:", "1. Thaw one vial of PBMCs in the water bath. Transfer immediately to 6mL of room temperature SCAIP media and mix gently using a wide-bore tip. Rinse t...
69,885
Stereotaxic rat brain surgery for Substantia Nigra targeting
4
dx.doi.org/10.17504/protocols.io.dm6gpj8zpgzp/v1
https://www.protocols.io/view/stereotaxic-rat-brain-surgery-for-substantia-nigra-cgg5tty6
miquel.vila
TITLE: Stereotaxic rat brain surgery for Substantia Nigra targeting AUTHORS: miquel.vila [DESCRIPTION] Stereotaxic rat brain surgery [STEPS] 1. Anesthetize rat with isofluorane (5% induction phase, 2% maintenance phase) 2. Weight rat & shave the head 3. Inject meloxicam (0.5 mg/mL) and Baytril (5 mg/mL) 4. Place rat...
["Anesthetize rat with isofluorane (5% induction phase, 2% maintenance phase)", "Weight rat & shave the head", "Inject meloxicam (0.5 mg/mL) and Baytril (5 mg/mL)", "Place rat head gently with ears bar onto the stereotaxic equipment", "Scalpel + Scalpel holder = incision middle of rat head", "Turn vacuum on", "Place 4 ...
26,535
U Michigan - Spot urine collection
null
dx.doi.org/10.17504/protocols.io.56fg9bn
null
Jeff Hodgin
TITLE: U Michigan - Spot urine collection AUTHORS: Jeff Hodgin [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block">Collection of urine from experimental animals is a basic requirement to detect albumin/creatinine ratio for ...
["Create 6 evenly spaced holes in the lid of the container – 3 on top and 3 at bottom using an electric drill using 1/8” drill bit. Also drill 2 holes each on four sides of the container. Using a metal cutter cut out a 14.5 cm by 9.5 cm piece of stainless steel wire mesh to create a base for mouse that will fit inside ...
36,914
NADH-linked microtiter plate-based assay for measuring Rubisco activity & activation state – GAPDH-GlyPDH
1
dx.doi.org/10.17504/protocols.io.bgasjsee
https://www.protocols.io/view/nadh-linked-microtiter-plate-based-assay-for-measu-bgasjsee
Cristina Rodrigues Gabriel Sales, Anabela Silva, Elizabete Carmo-Silva
TITLE: NADH-linked microtiter plate-based assay for measuring Rubisco activity & activation state – GAPDH-GlyPDH AUTHORS: Cristina Rodrigues Gabriel Sales, Anabela Silva, Elizabete Carmo-Silva [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol uses five reactions to couple RuBP carboxyla...
["[REAGENTS & SOLUTIONS]\nREAGENTS & SOLUTIONS TO PREPARE BEFOREHAND\nPowder chemical stocks stored at -20°C: let warm up to room temperature on desiccant before opening container.Expensive chemicals purchased in very small amounts (mg), for which concentration in assay is not critical (e.g. in excess): trust quantity ...
86,046
Luminescent Conjugated Oligothiophenes (LCO) Staining
1
dx.doi.org/10.17504/protocols.io.x54v9pd7pg3e/v1
https://www.protocols.io/view/luminescent-conjugated-oligothiophenes-lco-stainin-cx96xr9e
Toby J Curless, Hemanth Ramesh Nelvagal, Zane Jaunmuktane
TITLE: Luminescent Conjugated Oligothiophenes (LCO) Staining AUTHORS: Toby J Curless, Hemanth Ramesh Nelvagal, Zane Jaunmuktane [DESCRIPTION] The protocol describes Luminescent Conjugated Oligothiophenes staining on brain sections. [STEPS] SECTION: Preparation 1. Generate tissue sections using standard microtome s...
["[Preparation] Generate tissue sections using standard microtome sectioning protocols.", "[Deparaffinisation and LCO Staining] De-paraffinise sections by 5 min washes in xylene (x2), 5 min 100 % ethanol (x2) and 5 min in DEPC H2O (x1).", "[Preparation] Heat tissue dry tissue sections for 60 min at 60 °C", "[Deparaffin...
104,399
Genotyping for specific genomic insertions with Cas12a ssDNA cleavage
0
dx.doi.org/10.17504/protocols.io.14egn6n2pl5d/v1
https://www.protocols.io/view/genotyping-for-specific-genomic-insertions-with-ca-dh7p39mn
David Booth
TITLE: Genotyping for specific genomic insertions with Cas12a ssDNA cleavage AUTHORS: David Booth [DESCRIPTION] Based on the DETECTR assay, this protocol screens for precise genomic insertions by programming a CRISPR RNA (crRNA) to direct LbCas12a to recognize an amplified genomic locus. This sensitive assay enables t...
["[Set-up Cas12a ssDNA cleavage assay] 1440 minafter transfection count the cell density using a haemocytometer", "[Set-up Cas12a ssDNA cleavage assay] Dilute cells to a density of 45 cells/ml in a 50 mLconical tube.", "[Extract DNA from cell populations] Once cells have grown to saturation, remove 12 µL from each wel...
80,536
Modified Nucleic Acid Extraction Protocol for Humic-Rich Permafrost Peat Samples
4
dx.doi.org/10.17504/protocols.io.yxmvm244bg3p/v1
https://www.protocols.io/view/modified-nucleic-acid-extraction-protocol-for-humi-csvywe7w
Yueh-Fen Li, Rhiannon Mondav, Isogenie 1 Project Coordinators, Virginia Rich
TITLE: Modified Nucleic Acid Extraction Protocol for Humic-Rich Permafrost Peat Samples AUTHORS: Yueh-Fen Li, Rhiannon Mondav, Isogenie 1 Project Coordinators, Virginia Rich [DESCRIPTION] Slight modifications to the manufacturer’s protocol (Qiagen #12966-10, now discontinued and replaced with #12988-10) enabled the su...
["[Total nucleic acids extraction] Preheat the water bath to 60 ˚C. If the phenol:chloroform:isoamyl alcohol (PCI) (25:24:1, pH8.0) is refrigerated, move from 4 ˚C to the fume food to warm to room temperature (RT). CAUTION: PCI is toxic and needs to be handled in the fume hood with proper PPEs. Handle with care.", "[T...
40,603
Vezina Lab Vibratome Sectioning
1
null
https://www.protocols.io/view/vezina-lab-vibratome-sectioning-bjv3kn8n
Chad Vezina
TITLE: Vezina Lab Vibratome Sectioning AUTHORS: Chad Vezina [STEPS] ?. [At least one day prior to starting] Make molds and vibratome blades ?. [At least one day prior to starting] Soak forceps & molds in RNase inhibitor soln ?. [At least one day prior to starting] Prep low-melt agarose ?. [Prepare sample tissues] Remo...
["[At least one day prior to starting]\nMake molds and vibratome blades", "[At least one day prior to starting]\nSoak forceps & molds in RNase inhibitor soln", "[At least one day prior to starting]\nPrep low-melt agarose", "[Prepare sample tissues]\nRemove dehydrated tissue from –20°C storage", "[Prepare sample tissues...
null
null
null
dx.doi.org/10.17504/protocols.io.smvec66
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
31,118
Total DNA extraction from plant tissue using CTAB method
null
dx.doi.org/10.17504/protocols.io.bamnic5e
null
Robert Auber
TITLE: Total DNA extraction from plant tissue using CTAB method AUTHORS: Robert Auber [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. Using a pre-chilled mortar and pestle, grind frozen plant tissue (1-5g grams) into a fine powder ?. Add 500 ul of pre-heated 2 x CTAB buffer (with beta-mercaptoethanol added...
["Using a pre-chilled mortar and pestle, grind frozen plant tissue (1-5g grams) into a fine powder", "Add 500 ul of pre-heated 2 x CTAB buffer (with beta-mercaptoethanol added) to the pelleted cells. Vortex and re-suspend by gently pipetting up and down.\n60 °C", "Incubate at 60 C for 30 min. Periodically, mix gently d...
69,583
Biochemical detection of aggregated Tau
4
dx.doi.org/10.17504/protocols.io.n92ldpbx8l5b/v1
https://www.protocols.io/view/biochemical-detection-of-aggregated-tau-cf7ptrmn
Itika Saha, F. Ulrich Hartl, Mark S. Hipp
TITLE: Biochemical detection of aggregated Tau AUTHORS: Itika Saha, F. Ulrich Hartl, Mark S. Hipp [DESCRIPTION] This protocol describes the detection of aggregated Tau from HEK293 cells stably expressing and propagating aggregates of Tau repeat domain fused to YFP (Sanders et al. Neuron, 2014; Saha et al, BioRxiv, 202...
["[Preparation of cell lysates] The protocol can be performed with freshly harvested cells or with frozen cell pellets. In either case, harvest cells by trypsinization and wash 1x with PBS before pelleting in 1.5 mL Eppendorf tubes.", "[Preparation of cell lysates] Thaw frozen cell pellets on ice for at least 10 min.",...
28,129
Protein gel sample preparation
null
dx.doi.org/10.17504/protocols.io.7p9hmr6
null
Cleo B.
TITLE: Protein gel sample preparation AUTHORS: Cleo B. [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Sample preparation for tricine SDS-page gels and polyacrylamide SDS-page gels. </div></div> [STEPS] ?. [sample preparation tricine SDS page] If you are unsure about the amount of protein present ...
["[sample preparation tricine SDS page]\nIf you are unsure about the amount of protein present in your sample, load multiple concentrations Add beta-mercaptoethanol to tricine sample buffer until a final concentration of 2% Dilute 1 part sample with 1 part sample buffer", "[sample preparation SDS page]\nIf you are uns...
null
null
null
dx.doi.org/10.17504/protocols.io.pw9dph6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>For cDNA synthesis and for amplify larger fragments of YFV genome. The PCR target is of the CprM/envelope region of YFV genome described by Jorge and colleagues (2017).</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
50,876
Power Analysis Apo Field work
3
null
https://www.protocols.io/view/power-analysis-apo-field-work-bvw4n7gw
Tom Little, Nick Colegrave
TITLE: Power Analysis Apo Field work AUTHORS: Tom Little, Nick Colegrave [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>We used </span><span style = "font-weight:bold;">Power analyses</span><span> to clarify our ability to detect the effects of stressors as main effects, or as interactions, w...
[]
85,896
In vitro depalmitoylation assay
4
dx.doi.org/10.17504/protocols.io.bp2l6x9bklqe/v1
https://www.protocols.io/view/in-vitro-depalmitoylation-assay-cx5gxq3w
wusj, schekman
TITLE: In vitro depalmitoylation assay AUTHORS: wusj, schekman [DESCRIPTION] This protocol describes the in vitro depalmitoylation assay of DNAJC5 [STEPS] SECTION: In vitro depalmitoylation assay 1. HEK293T, MDA-MB-231, or Hela cells were transfected with plasmids encoding DNAJC5 by lipofectamine 2000 followed the co...
["[In vitro depalmitoylation assay] HEK293T, MDA-MB-231, or Hela cells were transfected with plasmids encoding DNAJC5 by lipofectamine 2000 followed the commercial protocol by Thermo Fisher.", "[In vitro depalmitoylation assay] Cellular membranes were prepared as described in \"Membrane and cytosol fractionation protoc...
null
null
null
dx.doi.org/10.17504/protocols.io.e4bbgsn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>For routine protein assays, we recommend that researchers generate their own CB-X™ Tables or standard curves. This is a one time commitment to ensure user specific results and allows single tube assays to be performed each time as opposed to generating new calibration plots f...
[]
81,046
Spore production of the wheat stripe rust pathogens in control growth cabinet.
4
null
https://www.protocols.io/view/spore-production-of-the-wheat-stripe-rust-pathogen-ctdwwi7e
Ramawatar Nagar
TITLE: Spore production of the wheat stripe rust pathogens in control growth cabinet. AUTHORS: Ramawatar Nagar [DESCRIPTION] Rust are biotrophic pathogens that can only be grown on the living organism and can not be grown in a lab on artificial media. Hence, the only tissue available for biological experiments are spo...
["Spore Production", "For spore production we normally use 30 pots and sow around 10-15 seeds per pot. The seeds shouldn’t be sow very deep in the soil.", "Put them in control growth cabinet and add some fertilizer to the surface of each pot. After that don’t water the plants for 5 days.", "After 4-5days or when the se...
18,824
sohbet
null
dx.doi.org/10.17504/protocols.io.wmgfc3w
null
Sohbet Odalari
TITLE: sohbet AUTHORS: Sohbet Odalari [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Sıcak </span><a style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">sohbet odaları</span></a><span>, yetişkin sohbet ve </span><a style = "text-decoration:underline;color:blue;cu...
["sohbet"]
38,029
Viability Estimation of Islets for Distribution Using Inclusion and Exclusion Fluorescent Dyes (FDA/PI)
1
dx.doi.org/10.17504/protocols.io.bhdmj246
https://www.protocols.io/view/viability-estimation-of-islets-for-distribution-us-bhdmj246
Integrated Islet Distribution Program
TITLE: Viability Estimation of Islets for Distribution Using Inclusion and Exclusion Fluorescent Dyes (FDA/PI) AUTHORS: Integrated Islet Distribution Program [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This Standard Operating Procedure is adapted from the work of the '</span><span style = ...
["[Islet Assessment Preparation]\nAssemble all items described in the “Assay Set Up” section.", "[Islet Assessment Preparation]\nPrepare for Fluorescent Microscopy and Image Recording: Refer to the Microscope and camera using manufacture instructions and settings.", "[Islet Assessment Preparation]\nPrepare [IIDP Islet ...
35,827
Detection of Sars-Cov2 Using Droplet Digital PCR
null
dx.doi.org/10.17504/protocols.io.be8tjhwn
https://www.protocols.io/view/detection-of-sars-cov2-using-droplet-digital-pcr-be8tjhwn
Joseph Patterson, Allyson Cole-Strauss, John Beck, Caryl Sortwell, Jack Lipton
TITLE: Detection of Sars-Cov2 Using Droplet Digital PCR AUTHORS: Joseph Patterson, Allyson Cole-Strauss, John Beck, Caryl Sortwell, Jack Lipton [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Assay sensitivity is an important part of any lab test, this becomes a more critical issue when dealing wit...
["[cDNA Synthesis]\nAdd 11 µL of Master mix to each PCR tube used. •4.4 µL iScript Reverse Transcription Supermix for RT-qPCR (Bio-Rad 1708841). •6.6 µL DNase/RNase free water.", "[cDNA Synthesis]\nAdd 11 µL of sample/diluted RNA to each tube (identify optimal dilution prior to performing the real experiment).", "[cDNA...
100,146
Semi-Automated Dual Fluorescence In-Situ Hybridization (d-FISH) Procedure
0
dx.doi.org/10.17504/protocols.io.x54v92dx4l3e/v1
https://www.protocols.io/view/semi-automated-dual-fluorescence-in-situ-hybridiza-dd2s28ee
Allen Institute
TITLE: Semi-Automated Dual Fluorescence In-Situ Hybridization (d-FISH) Procedure AUTHORS: Allen Institute [DESCRIPTION] This semi-automated protocol provides step by step instructions for performing Dual-Fluorescence In Situ Hybridization on tissue. [STEPS]
[]
40,799
ELISA for quantification of IL-35 in human serum.
6
dx.doi.org/10.17504/protocols.io.bj37kqrn
https://www.protocols.io/view/elisa-for-quantification-of-il-35-in-human-serum-bj37kqrn
Angel Justiz-Vaillant
TITLE: ELISA for quantification of IL-35 in human serum. AUTHORS: Angel Justiz-Vaillant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells....
["An anti-human IL-35 coating antibody is adsorbed onto the microwells by incubation overnight at 4°C with carbonate-bicarbonate buffer.", "Add 50 µl of human serum. Human IL-35 present in the serum sample binds to antibodies adsorbed into the microwells.", "The microplate is blocked with 3% non-fat milk-PBS buffer and...
84,324
High-Molecular-Weight SPRI-aided DNA extraction from Mimulus (Phrymaceae) leaf tissue
4
dx.doi.org/10.17504/protocols.io.bp2l6xn8rlqe/v2
https://www.protocols.click/view/high-molecular-weight-spri-aided-dna-extraction-fr-cwkcxcsw
Bolívar Aponte Rolón
TITLE: High-Molecular-Weight SPRI-aided DNA extraction from Mimulus (Phrymaceae) leaf tissue AUTHORS: Bolívar Aponte Rolón [DESCRIPTION] This protocol is adapted from Russo et al. 2023 methods for high-molecular weight DNA extraction. I modified the protocol to use reagents and incubation conditions used by the Ferris...
["[Sample preparation] Clean the biosafety cabinet, pipettes, tip boxes and any other instrument to be used with 70% EtOH, 10 % Bleach and 95 % EtOH. Surface sterilize all instruments in biosafety cabinet with UV light for 30 min.", "[Sample preparation] Quickly after placing tissue in tubes, place tube cap and close ...
50,434
Microfluidic Chip Production
1
null
https://www.protocols.io/view/microfluidic-chip-production-bvhan32e
Suresh Poovathingal, Florian De Rop, Stein Aerts
TITLE: Microfluidic Chip Production AUTHORS: Suresh Poovathingal, Florian De Rop, Stein Aerts [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for producing microfluidic chips used in HyDrop experiment.</div></div> [STEPS] ?. [Microfluidic Chip Preparation] Microfluidic device fabricationGu...
["[Microfluidic Chip Preparation]\nMicrofluidic device fabricationGuide to making microfluidic devices for bead production. The same method is used for the droplet encapsulation chips.", "[Microfluidic Chip Preparation]\nBake 4h @ 100 °C and store the finished chips with tape covering the inlets.", "[Microfluidic Chip ...
104,466
HIV-PULSE Wet-lab Protocol
0
dx.doi.org/10.17504/protocols.io.8epv5rby4g1b/v2
https://www.protocols.io/view/hiv-pulse-wet-lab-protocol-dh9s396e
Laurens Lambrechts, Sofie De Braekeleer, Basiel Cole, Linos Vandekerckhove
TITLE: HIV-PULSE Wet-lab Protocol AUTHORS: Laurens Lambrechts, Sofie De Braekeleer, Basiel Cole, Linos Vandekerckhove [DESCRIPTION] Protocols.io for HIV-PULSE assay, including an updated protocol from the work published in NAR. [BEFORE_START] The HIV-PULSE assay consists of six different PCR steps followed by sequenc...
["[Pre-amplification: PCR using PrimeSTAR GXL] Prepare the master mix according to this principle (if doing replicates, add DNA to master mix, mix well and then divide per sample to ensure ‘even’ input), beware to adapt the mix according to the required DNA input volume: \na. x µL DNA input \nb. 33- x µL NFW", "[Pre-am...
62,538
Keto Now [SCAM Or LEGIT] Supplement Official Website!!
3
dx.doi.org/10.17504/protocols.io.e6nvwkom7vmk/v1
https://www.protocols.io/view/keto-now-scam-or-legit-supplement-official-website-b9bir2ke
Keto Now
TITLE: Keto Now [SCAM Or LEGIT] Supplement Official Website!! AUTHORS: Keto Now [DESCRIPTION] https://www.ottawalife.com/article/keto-now-reviews-canada-avis-en-francais-does-keto-now-pills-price-worthy [STEPS]
[]
104,111
Isolation of brain infiltrating lymphocytes
0
dx.doi.org/10.17504/protocols.io.dm6gpzo68lzp/v2
https://www.protocols.io/view/isolation-of-brain-infiltrating-lymphocytes-dhwp37dn
Moustafa Nouh Elemeery, Salix Boulet, louis-eric.trudeau, Nathalie Labrecque
TITLE: Isolation of brain infiltrating lymphocytes AUTHORS: Moustafa Nouh Elemeery, Salix Boulet, louis-eric.trudeau, Nathalie Labrecque [DESCRIPTION] This protocol details the isolation of brain infiltrating lymphocytes. Splenic lymphocytes are screened to verify the presence/phenotype of CD8 in the periphery. [STE...
["[Infiltration procedure] Inject mice intravenously with CD45-FITC 3ug/mice (6 µL from vial/mice) for 3 min-5 min, then euthanized directly by decapitating head to stop blood circulation.", "[Infiltration procedure] Collect the brain for each experimental condition vs control.", "[Infiltration procedure] Dissociate br...
null
null
null
dx.doi.org/10.17504/protocols.io.fkabkse
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Non Neutrophil cells are depleted by incubating the sample with the biotin antibody cocktail followed by incubation with magnetic Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. The untouched cells are collected. These...
[]
105,302
Fixation and Immunostaining
0
null
https://www.protocols.io/view/fixation-and-immunostaining-di3w4gpe
Gist Croft, mskowronska Skowronska
TITLE: Fixation and Immunostaining AUTHORS: Gist Croft, mskowronska 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...
["[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 ...
90,648
Tissue Sectioning
1
dx.doi.org/10.17504/protocols.io.bp2l6xm8klqe/v1
https://www.protocols.io/view/tissue-sectioning-c4ryyv7w
Michael X. Henderson
TITLE: Tissue Sectioning AUTHORS: Michael X. Henderson [DESCRIPTION] This protocol details formalin-fixed paraffin-embedded tissue sectioning on a microtome for subsequent tissue staining. [STEPS] SECTION: Tissue Sectioning 1. Clean microtome blade with xylene (to remove paraffin) and then 100% ethanol (to dry). SECT...
["[Tissue Sectioning] Clean microtome blade with xylene (to remove paraffin) and then 100% ethanol (to dry).", "[Tissue Sectioning] Mount blade on microtome, and tighten screws to hold it in place.", "[Tissue Sectioning] Mount paraffin block in holder. Manually move the block near, but not touching the blade.", "[Tissu...
19,864
qPCR Identification Haplosporidium pinnae protocol
null
dx.doi.org/10.17504/protocols.io.xmyfk7w
null
Monserrat López-Sanmartín, Gaetano Catanese, Amalia Grau, Jose Maria Valencia, JRafa García-March, JINavas
TITLE: qPCR Identification Haplosporidium pinnae protocol AUTHORS: Monserrat López-Sanmartín, Gaetano Catanese, Amalia Grau, Jose Maria Valencia, JRafa García-March, JINavas [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Real time PCR to identify </span><span style = "font-style:italic;">Hapl...
[]
25,219
High Molecular Weight DNA Extraction from Recalcitrant Plant Species for Third Generation Sequencing
null
dx.doi.org/10.17504/protocols.io.4vbgw2n
null
Rachael Workman, Renee Fedak , Duncan Kilburn, Stephanie Hao, Kelvin Liu, Winston Timp
TITLE: High Molecular Weight DNA Extraction from Recalcitrant Plant Species for Third Generation Sequencing AUTHORS: Rachael Workman, Renee Fedak , Duncan Kilburn, Stephanie Hao, Kelvin Liu, Winston Timp [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left"...
["[Nuclei Isolation]\nGrind of tissue, preferably fresh or snap frozen, into fine powder in liquid nitrogen with a mortar and pestle. Immediately transfer ground tissue to capped 250 mL bottle containing NIB. Cap bottle and attach to end over end mixer, rotating at max speed for at .\n1 g\n10 ml\n4 °C", "[Nuclei Iso...
101,589
Structural and Functional Annotation of bee genome
0
dx.doi.org/10.17504/protocols.io.3byl49wwogo5/v1
https://www.protocols.io/view/structural-and-functional-annotation-of-bee-genome-dffv3jn6
Rafael Rodrigues Ferrari, Thiago Mafra Batista
TITLE: Structural and Functional Annotation of bee genome AUTHORS: Rafael Rodrigues Ferrari, Thiago Mafra Batista [DESCRIPTION] This protocol provides detailed, step-by-step instructions for students and researchers to annotate nuclear genomes, using the genome of the Asian bee Colletes collaris as an example. We begi...
["[IDENTIFICATION OF REPETITIVE REGIONS] Identification of repetitive regions\n\n****RepearModeler (on Kiko)****\n\n***Building a database***\n \n***Run RepeatModeler***\n \n****RepeatMasker (on kiko)****\n\n***Masking genome***\n\n#Use the -families.fa\n \n***Generating a summary***\n \n***Generating a .gff3 for use i...
64,440
Max Boost Keto Ultra Burn [BETTER OUT COME] Does It Really Work?
3
dx.doi.org/10.17504/protocols.io.5qpvobwmxl4o/v1
https://www.protocols.io/view/max-boost-keto-ultra-burn-better-out-come-does-it-ca6yshfw
H A
TITLE: Max Boost Keto Ultra Burn [BETTER OUT COME] Does It Really Work? AUTHORS: H A [DESCRIPTION] Also, your body will remain in ketosis consuming with smoldering heat fat assuming that you keep those ketone levels high. [STEPS]
[]
18,606
Modified HMW DNA Isolation from Stramenopiles with Agar Plugs
null
dx.doi.org/10.17504/protocols.io.wenfbde
null
Vincent Bielinski, Chris Dupont
TITLE: Modified HMW DNA Isolation from Stramenopiles with Agar Plugs AUTHORS: Vincent Bielinski, Chris Dupont [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is the adaptation of previously published methods for use in isolating intact high molecular weight DNA from stramenopiles for l...
["Add 0.5 g of LMP agarose to 25 mL TAE (2% final)", "Microwave for 30 secs or until solution begins to bubble", "Repeat heating steps as necessary (2-3x)", "Move flask to 50oC air incubator, swirling a few times to bring agarose into solution", "Thaw 50 mL conical vial with cell pellets on ice (cell pellet ~1.5g for t...
28,339
MojoSort™ Mouse CD4 Nanobeads Protocol
null
dx.doi.org/10.17504/protocols.io.7wthpen
null
Sam Li
TITLE: MojoSort™ Mouse CD4 Nanobeads Protocol AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Product description and procedure summary:</span></div><div class = "text-block">The cells targeted by the Nanobeads are either selected or depleted by incu...
["Prepare cells from your tissue of interest or blood without lysing erythrocytes.", "In the final wash of your sample preparation, resuspend the cells in MojoSort™ Buffer by adding up to 4 mL in a 5 mL (12 x 75 mm) polypropylene tube.Note: Keep MojoSort™ Buffer on ice throughout the procedure.", "Filter the cells with...
null
null
null
dx.doi.org/10.17504/protocols.io.i2gcgbw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The simple pipeline to examine the natural variation in the coding region of your favourite gene, that you either picked by GWAS or just working on for the entire life and would like to examine what else is there to it. The pipeline described in this protocol is based on the ...
[]
90,751
Elution of nasal lining fluid collected via nasosorption
1
dx.doi.org/10.17504/protocols.io.14egn3nn6l5d/v1
https://www.protocols.io/view/elution-of-nasal-lining-fluid-collected-via-nasoso-c4u7ywzn
Samuel Montgomery
TITLE: Elution of nasal lining fluid collected via nasosorption AUTHORS: Samuel Montgomery [DESCRIPTION] This procedure outlines the materials and processes for eluting nasal lining fluid collected using a Mucosal Diagnostics FX-i nasosorption device. This protocol has been successfully used to elute nasal lining flui...
["[Making elution buffer] Add 0.05 % volume of Tween20 to PBS in a sterile environment to reduce cross-contamination", "[Making elution buffer] Store at 4 °C until required", "[Elution of nasal lining fluid] Thaw nasosorption device in cryotube on ice", "[Elution of nasal lining fluid] Add 300 µL of elution buffer to ...
32,148
nCoV-2019 sequencing protocol
null
dx.doi.org/10.17504/protocols.io.bbmuik6w
null
Josh Quick
TITLE: nCoV-2019 sequencing protocol AUTHORS: Josh Quick [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">ARTIC amplicon sequencing protocol for MinION for nCoV-2019</div></div> [STEPS] ?. [cDNA preparation] Mix the following components in an 0.2mL 8-strip tube;Component ...
["[cDNA preparation]\nMix the following components in an 0.2mL 8-strip tube;Component Volume50µM random hexamers 10mM dNTPs mix (10mM each) Template RNA Total\n1 µl\n1 µl\n11 µl\n13 µl\nViral RNA input from a clinical sample sho...
28,178
The &#39;Three Peaks&#39; faecal DNA extraction method for long-read sequencing
null
dx.doi.org/10.17504/protocols.io.7rshm6e
null
Josh Quick
TITLE: The &#39;Three Peaks&#39; faecal DNA extraction method for long-read sequencing AUTHORS: Josh Quick [STEPS] ?. Add fresh stool or OMNIgene GUT to DNA/RNA Shield, vortex briefly and place on a tube rotator for at . 100 mg 50 µl 200 µl Centrifuge: 20 33 ?. Centrifuge at for and retain up to supernatant depe...
["Add fresh stool or OMNIgene GUT to DNA/RNA Shield, vortex briefly and place on a tube rotator for at .\n100 mg\n50 µl\n200 µl\nCentrifuge: 20 33", "Centrifuge at for and retain up to supernatant depending on size of pellet.\nCentrifuge: 5000 34\n200 µl", "Add PBS and resuspend material by pipetting up and down...
94,434
Tn5-Duplex-Sequencing (Tn5-Duplex-Seq) for low-input single-molecule variant detection
1
dx.doi.org/10.17504/protocols.io.6qpvr3nbzvmk/v1
https://www.protocols.io/view/tn5-duplex-sequencing-tn5-duplex-seq-for-low-input-c8gaztse
Diane Shao, Nazia Hilal, Sangita Choudhury
TITLE: Tn5-Duplex-Sequencing (Tn5-Duplex-Seq) for low-input single-molecule variant detection AUTHORS: Diane Shao, Nazia Hilal, Sangita Choudhury [DESCRIPTION] DNA mutations are the inevitable consequences of errors that arise during replication-repair of DNA damage as well as aging and disease progression. Because of...
["[RECIPE FOR MAKING IN-HOUSE REAGENTS] 2X Single Cell Lysis Buffer \n 40 millimolar (mM) \n 40 millimolar (mM) \n 0.3 % volume \n \n \n\n12X quenching solution\n 600 millimolar (mM) \n 90 millimolar (mM) 0.02 % volume \n\n \nADP1 and ADP2 Mix\n Reconstitute the 16 ADP1 and 16 ADP2 primers separately in l...
43,439
Lab Notebook Template
3
null
https://www.protocols.io/view/lab-notebook-template-bnnpmddn
TITLE: Lab Notebook Template AUTHORS: [STEPS]
[]
51,758
Agar bioplastic (simmered) Ag01
1
null
https://www.protocols.io/view/agar-bioplastic-simmered-ag01-bwsnpede
Clara Davis
TITLE: Agar bioplastic (simmered) Ag01 AUTHORS: Clara Davis [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Agar bioplastic (simmered) Ag01</div></div> [STEPS] ?. Mix all of the ingredients together in a pot in the amounts above, and stir. Agar , Water and Glycerol 12 g 400 mL 18 g ?. Keep mixing...
["Mix all of the ingredients together in a pot in the amounts above, and stir. Agar , Water and Glycerol\n12 g\n400 mL\n18 g", "Keep mixing until there are no clumps and it is as dispersed as it’s gong to get. Then heat the mixture to boiling point, and simmer for 15 - 20 min, stirring constantly.", "Scoop out excess...
83,417
Full 18S metabarcoding of environmental samples of various substrats with Minion Nanopore
4
dx.doi.org/10.17504/protocols.io.3byl4qe48vo5/v1
https://www.protocols.io/view/full-18s-metabarcoding-of-environmental-samples-of-cvpzw5p6
Quentin Blandenier, Nicholas Gibson, Alexander K. Tice, Robert E Jones, Erin P. Jones, Richard E. Baird, Brendan A. Zurweller, Matthew W. Brown
TITLE: Full 18S metabarcoding of environmental samples of various substrats with Minion Nanopore AUTHORS: Quentin Blandenier, Nicholas Gibson, Alexander K. Tice, Robert E Jones, Erin P. Jones, Richard E. Baird, Brendan A. Zurweller, Matthew W. Brown [DESCRIPTION] We describe here a flexible protocol for eDNA metabarc...
["[Sampling] Select a pristine area without anthropic disturbance and collect your sample of interest (e.g. ~0.5-1 kg of soil, ~50-200 g of dry leaves or bark, ~1-2 L of water) in a clean plastic bag or bottle. Register all valuable sample information such as the date, label of the sample, localization of the site, coo...
null
null
null
dx.doi.org/10.17504/protocols.io.c47yzm
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol is for phospho-Tyrosine western blotting (optimized for detecting phospho FMRP protein after IP)</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
87,747
Total Starch Enzymatic Digestion
1
null
https://www.protocols.io/view/total-starch-enzymatic-digestion-czxbx7in
Lynn Doran, Amanda P. De Souza
TITLE: Total Starch Enzymatic Digestion AUTHORS: Lynn Doran, Amanda P. De Souza [DESCRIPTION] Enzymatic digestion of total soluble starch to glucose in plant tissue extracts for preparation for quantification via the GOD-POD Method (NZYtech). [BEFORE_START] Extract and dry total starch pellet from plant tissue per E...
["Prepare fresh daily 120 U/mL α-amylase (Bacillus licheniformis) in MOPS buffer. 1 mL per sample will be needed. Initial concentration of α-amylase is 3000 U/mL. Use C1V1 = C2V2 to calculate the volume of α-amylase and MOPS buffer to use.", "Prepare fresh daily 30 U/mL amyloglucosidase (Aspergillus niger) in acetate...
81,385
Fresh Frozen / OCT Tissue RNA Quality Evaluation -- University of Minnesota TMCs (ThermoFisher Scientific, Invitrogen - Purelink)
1
dx.doi.org/10.17504/protocols.io.dm6gpjpz8gzp/v1
https://www.protocols.io/view/fresh-frozen-oct-tissue-rna-quality-evaluation-uni-ctqhwmt6
ThermoFisher Scientific, Laura J Niedernhofer, David A Bernlohr
TITLE: Fresh Frozen / OCT Tissue RNA Quality Evaluation -- University of Minnesota TMCs (ThermoFisher Scientific, Invitrogen - Purelink) AUTHORS: ThermoFisher Scientific, Laura J Niedernhofer, David A Bernlohr [DESCRIPTION] RNA quality evaluation of OCT (Optimal cutting temperature) or fresh frozen blocks/samples. Com...
["[Purelink RNA Mini Kit Manual]", "[Purelink RNA Mini Kit Manual] Product Information \nCatalog Number: 12183018A\nhttps://www.thermofisher.com/order/catalog/product/12183018A \n\nor \n\nCatalog Number: 12183020\nhttps://www.thermofisher.com/order/catalog/product/12183020", "[Purelink RNA Mini Kit Manual] Complete: \...
100,788
SeV standard and copyback genomes PCR Protocols
0
dx.doi.org/10.17504/protocols.io.5qpvok3j7l4o/v1
https://www.protocols.io/view/sev-standard-and-copyback-genomes-pcr-protocols-denu3dew
Carolina Lopez
TITLE: SeV standard and copyback genomes PCR Protocols AUTHORS: Carolina Lopez [DESCRIPTION] Protocols for the detection of Sendai Virus (SeV) standard and copy-back genomes by PCR [BEFORE_START] PRINCIPLES BEHIND THE PROCEDURE MUST BE UNDERSTOOD. PLEASE CONSULT WITH EXPERIENCED LAB MEMBER THE FIRST TIME YOU USE THIS...
["[Reverse Transcription (RT)] *Different RT kits and steps for cbVG and Genome\n 1/3) cbVG: Use SuperScript III Kit\n\nProcedure\n*RT reaction can be completed in a 10 µL reaction or a 20 µLreaction. The 10 µLreaction is preferred, if RNA concentrations allow, to help conserve reagents.\n 1. Start with 1 µg ...
99,883
Tissue Quality Evaluation for Brain Perfusion/Dissection Specimens
1
dx.doi.org/10.17504/protocols.io.kxygxeonwv8j/v3
https://www.protocols.io/view/tissue-quality-evaluation-for-brain-perfusion-diss-ddsj26cn
Allen Institute for Brain Science
TITLE: Tissue Quality Evaluation for Brain Perfusion/Dissection Specimens AUTHORS: Allen Institute for Brain Science [DESCRIPTION] This protocol provides guidance in the assessment of quality of tissues post perfusion and dissection and applies to all such procedures. Perfusion and dissection quality will be categoriz...
[]
47,996
Criteria for performing total knee arthroplasty without postoperative intensive care monitoring
1
dx.doi.org/10.17504/protocols.io.bs44ngyw
https://www.protocols.io/view/criteria-for-performing-total-knee-arthroplasty-wi-bs44ngyw
Fabricio Loures, João Henrique Reis, Marcelo de Oliveira Campos, Guilherme Morgado Runco, Luísa Doim Vieira Fonseca, Luciana Ferreira Xavier, Nelson Hiroyuki Miyabe Ooka, Liszt Palmeira de Oliveira
TITLE: Criteria for performing total knee arthroplasty without postoperative intensive care monitoring AUTHORS: Fabricio Loures, João Henrique Reis, Marcelo de Oliveira Campos, Guilherme Morgado Runco, Luísa Doim Vieira Fonseca, Luciana Ferreira Xavier, Nelson Hiroyuki Miyabe Ooka, Liszt Palmeira de Oliveira [DESCRIPT...
[]
64,163
DNA Extract wash from filter paper stored at room temperature
1
dx.doi.org/10.17504/protocols.io.kxygxp2nzl8j/v4
https://www.protocols.io/view/dna-extract-wash-from-filter-paper-stored-at-room-cawbsfan
Katie Izenour, Anwar Kalalah, Fayez Salib, Sarah Zohdy
TITLE: DNA Extract wash from filter paper stored at room temperature AUTHORS: Katie Izenour, Anwar Kalalah, Fayez Salib, Sarah Zohdy [DESCRIPTION] Preservation of samples requiring cold-chain storage is an often unavoidable challenge especially when doing laboratory work outside the western world. Samples are a pr...
["[Whole blood collection from animal] Using a sterile syringe and needle, collect 1-5 mL of whole blood from animal's forelimb. Immediately express the contents of the syringe into a new, clean EDTA tube and invert several times to mix.", "[Whole blood storage] IF EXTRACTING DNA ON THE SAME DAY AS BLOOD COLLECTION - ...
81,563
Methodology for DMPs analysis
1
dx.doi.org/10.17504/protocols.io.n2bvj87jpgk5/v1
https://www.protocols.click/view/methodology-for-dmps-analysis-ctv3wn8n
Sara Coppini, bianca.gualandi, Giulia Caldoni, Mario Marino, Silvio Peroni, francesca.masini
TITLE: Methodology for DMPs analysis AUTHORS: Sara Coppini, bianca.gualandi, Giulia Caldoni, Mario Marino, Silvio Peroni, francesca.masini [DESCRIPTION] All eyes on data: Unleashing the untapped potential of research at the University of Bologna Led by data stewards at the University of Bologna, this preliminary pro...
["[Data collection] Using the DMPs and GAs of European projects as input, we structured the table in which to collect data information with the following variables or fields and their meaning or accepted values:\nProject identifier (project_id): alphanumeric string to identify the project to which the described data be...
36,423
Qant-iT™ PicoGreen® dsDNA Quantification
null
dx.doi.org/10.17504/protocols.io.bftfjnjn
https://www.protocols.io/view/qant-it-picogreen-dsdna-quantification-bftfjnjn
Roey Angel, Eva Petrova
TITLE: Qant-iT™ PicoGreen® dsDNA Quantification AUTHORS: Roey Angel, Eva Petrova [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>The following protocol is intended for the quantification of double-stranded DNA using </span><a href="https://www.thermofisher.com/order/catalog/product/P7589?ICID=...
["[Prepare reaction]\nTake out all reagents from the fridge and bring them to room temperature.Take out the DNA samples from the freezer. DNA samples should be slowly thawed on ice.\nQuant-iT™PicoGreen® dsDNA reagent is dissolved in dimethylsulfoxide (DMSO), which freezes below 19 °C. The reagent must be completely tha...
37,130
DH5α bacteria transformation protocol
null
dx.doi.org/10.17504/protocols.io.bghijt4e
https://www.protocols.io/view/dh5-bacteria-transformation-protocol-bghijt4e
Yoan Coudert
TITLE: DH5α bacteria transformation protocol AUTHORS: Yoan Coudert [STEPS] ?. Thaw bacteria on ice ?. Dispense 50μl in a microtube ?. Add 1 to 5 μl of DNA, and carefully flick the tube to mix cells and DNA ?. Place the mixture on ice for 10 minutes ?. Heat shock at 42°C for 30 seconds ?. Place on ice for 5 minutes ?. ...
["Thaw bacteria on ice", "Dispense 50μl in a microtube", "Add 1 to 5 μl of DNA, and carefully flick the tube to mix cells and DNA", "Place the mixture on ice for 10 minutes", "Heat shock at 42°C for 30 seconds", "Place on ice for 5 minutes", "Pipette 450 μl of room temperature SOC (or LB) into the mixture", "Incubate a...
25,960
Collect of Collodarian (Rhizaria, Radiolaria) nuclei for genomic analyses
1
dx.doi.org/10.17504/protocols.io.5kgg4tw
https://www.protocols.io/view/collect-of-collodarian-rhizaria-radiolaria-nuclei-5kgg4tw
Estelle Bigeard, Loïc Pillet, John Burns, Fabrice Not
TITLE: Collect of Collodarian (Rhizaria, Radiolaria) nuclei for genomic analyses AUTHORS: Estelle Bigeard, Loïc Pillet, John Burns, Fabrice Not [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Collodaria are ubiquitous and abundant marine radiolarian (Rhizaria) protists (Biard</span><span styl...
["[Solutions]\nPreparation of Solutions", "[Solutions]\nSucrose 3M (M = 342.3g / mol)112.96g in 110ml water milliQStore at room temperature", "[Solutions]\nSpermine 0.1M (M = 202.34g / mol) Powder stored at 4 ° C. Weigh 40.5mg in a 15ml Falcon.then add 2ml water milliQ.Preparation instructions: This product is soluble...
71,690
Isoflurane Anesthesia Protocol
1
dx.doi.org/10.17504/protocols.io.36wgqj7m5vk5/v1
https://www.protocols.io/view/isoflurane-anesthesia-protocol-ch9it94e
Michaela Selby, Alicia Avelar
TITLE: Isoflurane Anesthesia Protocol AUTHORS: Michaela Selby, Alicia Avelar [DESCRIPTION] protocol describes how to use anesthesia machine with isoflurane to anesthetize a rat. [BEFORE_START] Handle rat beforehand to reduce stress Bring rat from vivarium (or wherever it is kept) to surgery room in a clean cage [G...
["[Setup] Make sure you have a surgery room booked", "[Setup] Some sort of table or surgical workbench to transfer rat to when fully anesthetized", "[Setup] Grab isoflurane cart and/or supplies and bring it to the surgery room", "[Setup] Place paper towels in the plastic container", "[Setup] Make sure the tubing is tig...
84,443
Induction of non-selective bulk autophagy
4
dx.doi.org/10.17504/protocols.io.4r3l228b3l1y/v1
https://www.protocols.io/view/induction-of-non-selective-bulk-autophagy-cwp3xdqn
Elias Adriaenssens
TITLE: Induction of non-selective bulk autophagy AUTHORS: Elias Adriaenssens [DESCRIPTION] This protocol describes how to induce bulk (non-selective) autophagy in HeLa cells through nutrient starvation. [STEPS] SECTION: Nutrient starvation experiments 1. To induce bulk autophagy, starve the cells by culturing them i...
["[Nutrient starvation experiments] To induce bulk autophagy, starve the cells by culturing them in Hank balanced salt medium\n(HBSS, Thermo Fisher) for the indicated time.", "[Nutrient starvation experiments] Collect cells by trypsinization, wash them in PBS and lyse them in RIPA buffer for 20 min on ice. Clear the sa...
null
null
null
dx.doi.org/10.17504/protocols.io.hh3b38n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Organisms used for DNA transformation experiments were tested for sensitivity to five antibiotics in two different concentrations. Cell culture growing in F2 media with 3% seawater was divided to fifteen 15ml tubes, i.e. three tubes for testing each of the antibiotics: antibi...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.kfbctin
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
53,458
5N HCl
4
dx.doi.org/10.17504/protocols.io.byfsptne
https://www.protocols.io/view/5n-hcl-byfsptne
Jacquelina.Woods
TITLE: 5N HCl AUTHORS: Jacquelina.Woods [DESCRIPTION] This method was developed at the FDA’s Center for Food Safety and Applied Nutrition for GenomeTrakr’s pandemic response project, monitoring SARS-CoV-2 variants in wastewater​​. Protocols developed for this project cover wastewater collection, concentration, RNA e...
["Slowly add 10.4 mL , or equivalent to the water.", "Measure 14.6 mL of deionized or ultrapure water into sterile container.", "Store at 2-8 Room temperature."]
17,119
Molecular testing of carrion flies for rabbit calicivirus detection
null
dx.doi.org/10.17504/protocols.io.ux7exrn
null
Robyn Hall
TITLE: Molecular testing of carrion flies for rabbit calicivirus detection AUTHORS: Robyn Hall [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Carrion fies are relatively easy and inexpensive to collect, systematic sampling networks can be established, and rabbit calicivirus can be detected in flies...
["[Trapping of flies]\nEnsure trap has been thoroughly decontaminated by soaking in 10% household bleach for at least 30 minutes, followed by rinsing with water. Place attractant in a specimen jar covered with a gauze swab to prevent flies coming into direct contact with the bait.Place trap in collection location for o...
null
null
null
dx.doi.org/10.17504/protocols.io.j4acqse
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Alveolar macrophages were obtained from the specimens of surgically resected lung tissue. Pieces of lung tissues (~10-30 g) obtained from lung parts resected from patients with treatment-refractory pulmonary tuberculosis. Lung tissue specimens were cut into small pieces and r...
[]
99,787
Genomic DNA Extraction from Sorted Cells
0
dx.doi.org/10.17504/protocols.io.n2bvjn88wgk5/v1
https://www.protocols.io/view/genomic-dna-extraction-from-sorted-cells-ddpj25kn
Raining Wang, Melinda Wheelock
TITLE: Genomic DNA Extraction from Sorted Cells AUTHORS: Raining Wang, Melinda Wheelock [DESCRIPTION] This protocol details the extraction of genomic DNA from cells. [GUIDELINES] The DNeasy columns cannot take more than 5 x 106 cells per column. Larger pellets can be split into multiple extraction reactions, then com...
["[Extraction of genomic DNA from cells] Prepare a lysis master mix of the following reagents and mix by vortexing briefly:", "[Extraction of genomic DNA from cells] Aliquot 322 µL of this master mix into labeled 1.7-mL microcentrifuge tubes.", "[Extraction of genomic DNA from cells] Resuspend the pellets in 100 µL DPB...
64,043
prueba
5
dx.doi.org/10.17504/protocols.io.4r3l2opo3v1y/v1
https://www.protocols.io/view/prueba-casjsecn
WILSON DAVID CAMPOS ROJAS
TITLE: prueba AUTHORS: WILSON DAVID CAMPOS ROJAS [DESCRIPTION] prueba [STEPS] 1. abrir a puerta 2. cerrar la puerta 3. echar llave
["abrir a puerta", "cerrar la puerta", "echar llave"]
null
null
null
dx.doi.org/10.17504/protocols.io.tqfemtn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Calcium imaging - recording the calcium dynamics of cultures using fluorescent calcium dye, fluorescence microscopy and a camera. [STEPS] SECTION: Calcium imaging set up ?. SECTION: Wash culture before insertion of calcium dye ?. SECTION: Insertion of calcium dye ?. SECTION:...
["[Calcium imaging set up] {\"blocks\":[{\"key\":\"7pnu9\",\"text\":\"Prepare the set up as follows:\",\"type\":\"unstyled\",\"depth\":0,\"inlineStyleRanges\":[],\"entityRanges\":[],\"data\":[]},{\"key\":\"ccdeq\",\"text\":\"Perform time lapse data at 2x2 binning mode with a resolution of 500x502 and 51.948 frames per ...
null
null
null
dx.doi.org/10.17504/protocols.io.gysbxwe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>SDS-PAGE using gels with low crosslinking of acrylamide and bisacrylamide. This protocol can be used to separate phosphorylation forms of KaiC proteins.</p> [GUIDELINES] <p><strong>references:</strong></p> <p> </p> <p><strong>1. SDS-PAGE</strong> modified from </p> <p> </p> ...
[]
88,713
DToL Tissue and Blood Sampling Standard Operating Procedure: Chordata: Vertebrata: Aves
1
dx.doi.org/10.17504/protocols.io.dm6gp3yedvzp/v1
https://www.protocols.io/view/dtol-tissue-and-blood-sampling-standard-operating-c2vhye36
Inez Januszczak, Nancy Holroyd
TITLE: DToL Tissue and Blood Sampling Standard Operating Procedure: Chordata: Vertebrata: Aves AUTHORS: Inez Januszczak, Nancy Holroyd [DESCRIPTION] This Standard Operating Procedure (SOP) contains guidance on how to sample tissue and blood from bird specimens submitted to the Darwin Tree of Life (DToL) project. Plea...
[]
93,741
TET buffer
3
dx.doi.org/10.17504/protocols.io.3byl4qy8ovo5/v1
https://www.protocols.io/view/tet-buffer-c7smznc6
Sarah Nagel, Anna Schmidt, Matthias Meyer
TITLE: TET buffer AUTHORS: Sarah Nagel, Anna Schmidt, Matthias Meyer [DESCRIPTION] TET buffer (10 mM Tris-HCl, 1 mM EDTA, 0.05% Tween-20, pH 8.0) is used in various steps of sample preparation by the Ancient DNA Core Unit of the MPI-EVA. [STEPS]
[]
81,576
Testing2
1
null
https://www.protocols.io/view/testing2-ctwgwpbw
Sebastian Vincent
TITLE: Testing2 AUTHORS: Sebastian Vincent [DESCRIPTION] est [STEPS] SECTION: Test 1. cry for 30 s 344 °C
["[Test] cry for 30 s 344 °C"]
53,044
Sampling leaf tissue for analysis of NPQ Relaxation using Technologica Chlorophyll Fluorescence Imager Data.
4
dx.doi.org/10.17504/protocols.io.bx2upqew
https://www.protocols.io/view/sampling-leaf-tissue-for-analysis-of-npq-relaxatio-bx2upqew
Lynn Doran, gotarkar
TITLE: Sampling leaf tissue for analysis of NPQ Relaxation using Technologica Chlorophyll Fluorescence Imager Data. AUTHORS: Lynn Doran, gotarkar [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Sampling leaf tissue for analysis of NPQ Relaxation using </span><a href="https://www.protocols....
["[Leaf Tissue Sampling]\nJust cover the bottom of a each well in a 24 well plate with water.", "[Leaf Tissue Sampling]\nUsing a #2 Humboldt cork borer, take leaf tissue samples from each plot for each technical replicate. Hold the leaf flat against the rubber stopper, press the cork borer through the leaf into the st...
61,244
70% Ethanol Fixation
4
null
https://www.protocols.io/view/70-ethanol-fixation-b724rqgw
Arnold Federico
TITLE: 70% Ethanol Fixation AUTHORS: Arnold Federico [DESCRIPTION] This protocol is specifically written for fixing/permeabilizing suspension cell culture in a final volume of 1mL. For DNA content analysis, such as for cell cycle analysis, a precipitating fixative such as ethanol is preferred over formaldehyde fixati...
["[Protocol] Transfer 1 * 106 to 5 * 106 suspended cells to appropriate centrifuge tube and pellet by spinning at 300g for 5 minutes", "[Protocol] Discard supernatant without disturbing pellet. Resuspend and wash cells in 1mL 1X PBS with gentle pipetting. Spin down cells again at 300g for 5 minutes.", "[Protocol] While...
86,511
TMA TNP Multiplex IHC Image Processing
5
dx.doi.org/10.17504/protocols.io.n92ldmmznl5b/v1
https://www.protocols.io/view/tma-tnp-multiplex-ihc-image-processing-cyqpxvvn
Shamilene Sivagnanam, Courtney Betts, Lisa Coussens
TITLE: TMA TNP Multiplex IHC Image Processing AUTHORS: Shamilene Sivagnanam, Courtney Betts, Lisa Coussens [DESCRIPTION] This specific protocol is for the TMA TNP cases. Understanding immune complexity within the tumor microenvironment provides valuable insight to tumor-immune composition, spatial interactions, and t...
["[File prep and Folder Structure] FOLDER AND FILE NAMING CONVENTION\n\nMake sure folder and file naming convention is consistent throughout the entire study. Filenaming convention should be used when saving images during acquisition.", "[File prep and Folder Structure] SET UP FOLDER STRUCTURE\n\nThe pipeline is optima...
null
null
null
dx.doi.org/10.17504/protocols.io.f35bqq6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
32,730
Comparative Efficacy of Antidepressants for Symptoms Remission of Gastroesophageal Reflux: A Bayesian Network Meta-Analysis of Randomized Controlled Trails (protocol)
null
dx.doi.org/10.17504/protocols.io.bb72irqe
null
Xiaobei Si, Deying Bi, Linyu Huo, Yu Lan, Shuo Zhang
TITLE: Comparative Efficacy of Antidepressants for Symptoms Remission of Gastroesophageal Reflux: A Bayesian Network Meta-Analysis of Randomized Controlled Trails (protocol) AUTHORS: Xiaobei Si, Deying Bi, Linyu Huo, Yu Lan, Shuo Zhang [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "f...
[]
51,428
Generation of Stable STING-GFP cells using retrovirus
1
dx.doi.org/10.17504/protocols.io.5jyl85xp7l2w/v1
https://www.protocols.io/view/generation-of-stable-sting-gfp-cells-using-retrovi-bwgcpbsw
Will Hancock-Cerutti, Pietro De Camilli
TITLE: Generation of Stable STING-GFP cells using retrovirus AUTHORS: Will Hancock-Cerutti, Pietro De Camilli [DESCRIPTION] This method describes the generation of HeLa cells stably expressing STING-GFP using retroviral transduction in order to study the localization of STING under different conditions. [STEPS] SECTI...
["[Cloning of pMX-STING-GFP retroviral vector] Amplify the coding sequence for human STING using PrimeSTAR GXL DNA polymerase (Takara Bio) according to manufacturer protocol. Primers include a XhoI restriction site at the 5’ end and a SacII restriction site at the 3’ end.", "[Cloning of pMX-STING-GFP retroviral vector]...
32,862
Isolation of cells from the epithelial layer of frozen human intestinal biopsies
null
dx.doi.org/10.17504/protocols.io.bcb6isre
null
Oxford HCA, Nicholas Provine, Michael FitzPatrick, Sophie Irwin
TITLE: Isolation of cells from the epithelial layer of frozen human intestinal biopsies AUTHORS: Oxford HCA, Nicholas Provine, Michael FitzPatrick, Sophie Irwin [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for the isolation of the epithelial cells (and associated immune cells wit...
["[Warming media]\nPer sample: pre-warm 10ml of Transport medium in the waterbath.", "[Warming media]\nPer sample: pre-warm 20ml of chelation medium in the waterbath.", "[Collection/Wash]\nPartially thaw frozen biopsies in 37°C waterbath until only a little ice remains.", "[Collection/Wash]\nDropwise add 1ml of pre-war...
46,969
PDI variants expression and purification protocol
4
dx.doi.org/10.17504/protocols.io.br4zm8x6
https://www.protocols.io/view/pdi-variants-expression-and-purification-protocol-br4zm8x6
Shihui Guo
TITLE: PDI variants expression and purification protocol AUTHORS: Shihui Guo [STEPS] ?. Human PDI was clone in the pT7-FLAG-SBP-1vector (sigma), designed for expression in E.coli BL21 (DE3) T1 cells (sigma). ?. Inoculate 10-ml Transformed E.coli BL21 (DE3) T1 cells on the medium containing ampicillin. ?. Grow the cult...
["Human PDI was clone in the pT7-FLAG-SBP-1vector (sigma), designed for expression in E.coli BL21 (DE3) T1 cells (sigma).", "Inoculate 10-ml Transformed E.coli BL21 (DE3) T1 cells on the medium containing ampicillin.", "Grow the culture O/N at 37degree.", "Transfer 10ml cell culture to 1Liter LB medium (100μg/ml Amp) ...
94,245
Protocol2_In vitro transcription.pdf
1
dx.doi.org/10.17504/protocols.io.ewov1qwxpgr2/v1
https://www.protocols.io/view/protocol2-in-vitro-transcription-pdf-c8adzsa6
angelica.gonzalez
TITLE: Protocol2_In vitro transcription.pdf AUTHORS: angelica.gonzalez [DESCRIPTION] In vitro transcription [STEPS]
[]
47,187
LID: imageanalyzer
5
dx.doi.org/10.17504/protocols.io.n2bvjxdkxlk5/v1
https://www.protocols.io/view/lid-imageanalyzer-bsbtnann
Saurin B Parikh
TITLE: LID: imageanalyzer AUTHORS: Saurin B Parikh [DESCRIPTION] LI Detector's imageanalyzer function makes the use of the MATLAB Colony Analyzer Toolkit to generate colony size estimations from image files. [STEPS] SECTION: LI Detector Analytical Pipeline 3. Execute Step 1 to 7 of the LI Detector Analytical Pipeline...
["[LI Detector Analytical Pipeline] Execute Step 1 to 7 of the LI Detector Analytical Pipeline\n\n\nNote: The order of the plates in the upscale pattern from Step 6 of the pipeline should be the same as the order of images within the folders.", "[Photo Ethics] Photos should be organized as Experiment > Arm > Stage > Ho...
68,583
Protocol for a nationwide systematic review of the Greek literature on child and adolescent mental health
1
dx.doi.org/10.17504/protocols.io.n92ldzo3nv5b/v1
https://www.protocols.io/view/protocol-for-a-nationwide-systematic-review-of-the-ce8fthtn
lauromarchionatti, Anastasia Koumoula, Arthur Caye, Vasiliki Eirini Karagiorga, Julia Luiza Schafer, Giovanni Abrahão Salum Júnior
TITLE: Protocol for a nationwide systematic review of the Greek literature on child and adolescent mental health AUTHORS: lauromarchionatti, Anastasia Koumoula, Arthur Caye, Vasiliki Eirini Karagiorga, Julia Luiza Schafer, Giovanni Abrahão Salum Júnior [DESCRIPTION] This is a landscape analysis of scientific literatur...
["[Protocol for a nationwide systematic review of the Greek literature on child and adolescent mental health] Protocol for a nationwide systematic review of the Greek literature on child and adolescent mental health\n\nAuthors and Affiliations\nAnastasia Koumoula1, Lauro Estivalete Marchionatti1,2,3 MD, Arthur Caye1,2,...
93,442
Multiplexed TMTpro proteomic sample preparation of whole cell HeLa lysates from various growth conditions ± FAC
4
dx.doi.org/10.17504/protocols.io.36wgq3kjolk5/v1
https://www.protocols.io/view/multiplexed-tmtpro-proteomic-sample-preparation-of-c7hazj2e
Felix Kraus, J. Wade Harper
TITLE: Multiplexed TMTpro proteomic sample preparation of whole cell HeLa lysates from various growth conditions ± FAC AUTHORS: Felix Kraus, J. Wade Harper [DESCRIPTION] The analysis of relative protein abundance has emerged as an important tool in cell biology. Typically, it is possible to quantify >8000 proteins un...
["[Harvest, precipitation and digestion] For whole proteome analysis, 50 µg is required for each replicate. Lyse cells in lysis buffer and pass them through a 21G needle 10 times. Alternatively, lyse cells by\nsonication as per manufactures instructions.", "[Harvest, precipitation and digestion] Centrifugate suspensio...
null
null
null
dx.doi.org/10.17504/protocols.io.fi3bkgn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>Experiment purpose is to monitor the time-course of a large-scale infection of host cyanobacteria by phage under variable media conditions and obtain samples for proteomic and transcriptomic analysis.</strong></p> <p> </p> <p><strong>15 Hourly Timepoints: 0, 1, 2, 3, ...
[]
84,091
In vitro kinase assay
4
dx.doi.org/10.17504/protocols.io.4r3l225xjl1y/v1
https://www.protocols.io/view/in-vitro-kinase-assay-cwc3xayn
Elias Adriaenssens
TITLE: In vitro kinase assay AUTHORS: Elias Adriaenssens [DESCRIPTION] This protocol describes in vitro kinase assay. [STEPS] SECTION: In vitro kinase assay 1. Mix recombinant proteins TBK1 or ULK1-complex (composed of ULK1, FIP200, ATG13, and ATG101) and NAP1 in kinase buffer. SECTION: In vitro kinase assay 2. Use t...
["[In vitro kinase assay] Mix recombinant proteins TBK1 or ULK1-complex (composed of ULK1, FIP200, ATG13, and ATG101) and NAP1 in kinase buffer.", "[In vitro kinase assay] Use the kinases at 50 nanomolar (nM) and mix with 250 nanomolar (nM) NAP1.", "[In vitro kinase assay] Start the kinase reactions by the adding 2x AT...
93,404
Cortical spheroid differentiation
4
dx.doi.org/10.17504/protocols.io.5jyl8po57g2w/v1
https://www.protocols.io/view/cortical-spheroid-differentiation-c7f4zjqw
Annika Martin, Hanqin Li, Dirk Hockemeyer
TITLE: Cortical spheroid differentiation AUTHORS: Annika Martin, Hanqin Li, Dirk Hockemeyer [DESCRIPTION] This protocol describes the procedure of differentiating hPSCs into early cortical spheroids following an adaptation of a published protocol (Yoon et al 2019, Paşca et al. 2015). Protocol overview A. Media Formul...
["[Media Formulations] hESC KSR Media (500 ml)\n 400 ml DMEM/F12\n 100 ml KSR\n 5 ml Pen-strep\n 5 ml NEAA\n 5 ml Glutamax\n 5 mL HEPES buffer\n\nhESC Wash Media (500 ml)\n 500 ml DMEM\n 25mL ml Calf Serum\n 5 ml Pen-strep\n\nhCS media (500 ml)\n 500 ml Neurobasal media\n 10 ...
16,336
Protocol for use with NEBNext® Small RNA Library Prep Set for Illumina® (E7300, E7580, E7560, E7330)
null
dx.doi.org/10.17504/protocols.io.t7qermw
null
New England Biolabs
TITLE: Protocol for use with NEBNext® Small RNA Library Prep Set for Illumina® (E7300, E7580, E7560, E7330) AUTHORS: New England Biolabs [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The NEBNext Multiplex Small RNA Library Prep Set for Illumina contains the adaptors, primers, enzymes and buffers ...
["[Ligate the 3´ SR Adaptor]\nMix the following components in a sterile nuclease-free PCR tube. It is ok to premix the reagents. Use immediately. AB1Input RNA1-6 µl2(green) 3 ́ SR Adaptor for Illumina1 µl3Nuclease-Free Watervariable4Total volume7 µl\nFor total RNA inputs of 100 ng, dilute the (green) 3´ SR Adaptor for...
59,879
Transfection for Recombinant Antibodies
4
dx.doi.org/10.17504/protocols.io.q26g741oqgwz/v1
https://www.protocols.io/view/transfection-for-recombinant-antibodies-b6qfrdtn
Addgene The Nonprofit Plasmid Repository
TITLE: Transfection for Recombinant Antibodies AUTHORS: Addgene The Nonprofit Plasmid Repository [DESCRIPTION] This protocol describes how to transfect suspension HEK293 cells with recombinant antibody plasmids using Polyethylenimine Max as a transfection reagent. After transfection and expression, the recombinant ant...
["[Seeding cells] One day prior to transfecting, seed a 108 mL of culture of cells at a density of 0.9*10^6 cells/mL in a 500 mL vented flask.", "[Seeding cells] Check cell density and viability:", "[Seeding cells] Transfer 108 mL of BCD TFX media into each of a 500 mL vented flask.", "[Seeding cells] Cap the flask and...
38,414
Hypertonic saline nasal irrigation and gargling for suspected or confirmed COVID-19: pragmatic web-based Bayesian adaptive randomised controlled trial (ELVIS COVID-19)
1
dx.doi.org/10.17504/protocols.io.bhrnj55e
https://www.protocols.io/view/hypertonic-saline-nasal-irrigation-and-gargling-fo-bhrnj55e
Aziz Sheikh, Sandeep Ramalingam, John Norrie, Emma Ward, Catriona Graham
TITLE: Hypertonic saline nasal irrigation and gargling for suspected or confirmed COVID-19: pragmatic web-based Bayesian adaptive randomised controlled trial (ELVIS COVID-19) AUTHORS: Aziz Sheikh, Sandeep Ramalingam, John Norrie, Emma Ward, Catriona Graham [DESCRIPTION] <div class = "text-blocks"><div class = "text-bl...
[]
84,860
E. coli recombineering protocol for gene deletions (SW102 prophage protocol)
4
dx.doi.org/10.17504/protocols.io.yxmvm3ke6l3p/v1
https://www.protocols.io/view/e-coli-recombineering-protocol-for-gene-deletions-cw44xgyw
is Sparrow
TITLE: E. coli recombineering protocol for gene deletions (SW102 prophage protocol) AUTHORS: is Sparrow [DESCRIPTION] This protocol is adapted from Thomason et al., 2023 https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10037674/ It is written for using the lambda red system (prophage) to make basic gene deletions in E. ...
["[Night before] The night before the experiment, incubate SW102 in liquid LB + tetracycline to grow at 30 °C at an appropriate rotation speed. \n\nNote: It is wise to check the temperature of your incubator with a mercury thermometer and calibrate as necessary, as temperatures above 32ºC cause leaky expression of prop...
26,537
U Michigan - Western Blot
null
dx.doi.org/10.17504/protocols.io.56hg9b6
null
Jeff Hodgin
TITLE: U Michigan - Western Blot AUTHORS: Jeff Hodgin [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span><span style = "font-weight:bold;"> </span></div><div class = "text-block">Immunoblotting (also called Western Blotting) is a technique to separate an...
["Wash the plates and wipe the surface with ethanol and dry plates.", "Assemble casting frames and glass plate. Put small glass plate on the top of big glass plate and fit whole thing into the casting stand with small plate outside. Close casting frame door, fill the space between two glass plates with H²O to check for...
38,554
FCMPASS - Performing and reporting calibration
5
dx.doi.org/10.17504/protocols.io.bhv2j68e
https://www.protocols.io/view/fcmpass-performing-and-reporting-calibration-bhv2j68e
Joshua Welsh, Jennifer Jones
TITLE: FCMPASS - Performing and reporting calibration AUTHORS: Joshua Welsh, Jennifer Jones [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol outlines the steps required to calculate the calibration parameters and convert fcs files using the FCMPASS software. This is one of a number of p...
["Upon completing fluorescence and/or light scatter calibration steps click the ‘Calibrate’ button.", "The FCMPASS software will now perform fluorescence and light scatter calibration. An FCMPASS export folder will be created in the directory from which the fcs files were imported. This folder will contain calibrated f...
46,460
PCR for SARS-CoV-2
3
dx.doi.org/10.17504/protocols.io.brk4m4yw
https://www.protocols.io/view/pcr-for-sars-cov-2-brk4m4yw
Leonardo Caserta
TITLE: PCR for SARS-CoV-2 AUTHORS: Leonardo Caserta [STEPS]
[]
82,154
7.1: Taxon Group: Crustacea - Decapoda
1
dx.doi.org/10.17504/protocols.io.261ge3y8ol47/v1
https://www.protocols.io/view/7-1-taxon-group-crustacea-decapoda-cugiwtue
Chris Fletcher, Paul Clark, Miranda Lowe, Lauren Hughes, Inez Januszczak
TITLE: 7.1: Taxon Group: Crustacea - Decapoda AUTHORS: Chris Fletcher, Paul Clark, Miranda Lowe, Lauren Hughes, Inez Januszczak [DESCRIPTION] This is part of the collection "DToL Taxon-specific Standard Operating Procedures (SOPs) for Marine Metazoa", lead by the Other Metazoa Working Group. The SOP collection contai...
[]
72,246
Agrobacterium-mediated transformation of the chytrid fungus Spizellomyces punctatus (Sp)
2
dx.doi.org/10.17504/protocols.io.x54v9dd1pg3e/v1
https://www.protocols.io/view/agrobacterium-mediated-transformation-of-the-chytr-ciswuefe
Sarah M Prostak, Edgar M Medina, Erik Kalinka, Lillian Fritz-Laylin
TITLE: Agrobacterium-mediated transformation of the chytrid fungus Spizellomyces punctatus (Sp) AUTHORS: Sarah M Prostak, Edgar M Medina, Erik Kalinka, Lillian Fritz-Laylin [DESCRIPTION] This is a collection of protocols for Agrobacterium-mediated transformation of the chytrid fungus Spizellomyces punctatus. [GUIDELI...
[]
98,105
Mechanical liver dissociation
0
dx.doi.org/10.17504/protocols.io.8epv5r1r5g1b/v1
https://www.protocols.io/view/mechanical-liver-dissociation-db2z2qf6
Dorien De Pooter, Ben De Clerck, Koen Dockx, Domenica De Santis, Sarah Sauviller, Pascale Dehertogh, Matthias Beyens, Isabelle Bergiers, Isabel Nájera, Ellen Van Gulck, Nádia Conceição-Neto, Wim Pierson
TITLE: Mechanical liver dissociation AUTHORS: Dorien De Pooter, Ben De Clerck, Koen Dockx, Domenica De Santis, Sarah Sauviller, Pascale Dehertogh, Matthias Beyens, Isabelle Bergiers, Isabel Nájera, Ellen Van Gulck, Nádia Conceição-Neto, Wim Pierson [DESCRIPTION] This protocol details the mechanical dissociation of liv...
["[Reagent preparation] Add 55 mL of FCS to a bottle of 500 mL of RPMI1640 with L-Glutamine.", "[Reagent preparation] Prepare GentleMACS C-tubes by filling them with 9 mL RPMI 1640 medium + 10% FCS and store the tubes at 4 °C.", "[Reagent preparation] Fill the syringe with 20 mLPBS and connect a 23G needle to the syrin...