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21,002
UC Davis - Lipoprotein profiling and partical size
null
dx.doi.org/10.17504/protocols.io.yrifv4e
null
Peter Havel
TITLE: UC Davis - Lipoprotein profiling and partical size AUTHORS: Peter Havel [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">Lipoprotein profiling and Lipoprotein size will be provided for CM, VLDL, LDL and HDL as ...
["The sample is loaded into a gel permeation column specifically designed for separating lipoprotein components. Then, lipoprotein is eluted from the column in order of larger particles to smaller ones.", "Lipoprotein fractionated by particle size is fed into reaction coils, and the degraded products after reacting wit...
81,743
Do, Q. B. &amp; Nebuloni, F. et al. (2023) A fluid walled microfluidic platform for human neuronal microcircuits and axotomy.
2
dx.doi.org/10.17504/protocols.io.36wgqjwwxvk5/v1
https://www.protocols.io/view/do-q-b-amp-nebuloni-f-et-al-2023-a-fluid-walled-mi-ct3pwqmn
Quyen Do, Federico Nebuloni, Richard Wade-Martins
TITLE: Do, Q. B. &amp; Nebuloni, F. et al. (2023) A fluid walled microfluidic platform for human neuronal microcircuits and axotomy. AUTHORS: Quyen Do, Federico Nebuloni, Richard Wade-Martins [DESCRIPTION] This collection contains six protocols detailing methods used in Do, Q. B. & Nebuloni, F. et al. (2023) A fluid w...
[]
53,868
Preparation of electrocompetent cells
4
dx.doi.org/10.17504/protocols.io.byukpwuw
https://www.protocols.io/view/preparation-of-electrocompetent-cells-byukpwuw
Shuning Guo
TITLE: Preparation of electrocompetent cells AUTHORS: Shuning Guo [DESCRIPTION] This protocol is used to prepare electrocompetent cells with high transformation efficiency. [BEFORE_START] All the medium and containers used in the protocol should be sterilized by high temperature autoclave. All the steps that expos...
["Line the E. coli strain on a solid LB medium without resistance and culture at 37℃ for 12h.", "Pick monoclonal cells from the medium and culture them in 5 ml LB medium at 37℃ for 12h.", "Inoculate 100 ml of LB medium with 1% volume of E. coli culture from step 2.", "Grow the cells at 37℃ shaking at 200 rpm to an OD60...
22,833
Measurement of duodenal motility using implanted strain gauges
null
dx.doi.org/10.17504/protocols.io.2irgcd6
null
Terry Powley, Zhenjun Tan, Matthew Ward
TITLE: Measurement of duodenal motility using implanted strain gauges AUTHORS: Terry Powley, Zhenjun Tan, Matthew Ward [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes a process for the measurement of electrical stimulation-induced effects on duodenal motility in young adult S...
["[Animals]\nTwo‐to four‐month‐old male rats were housed in vented rack cages in an Association for Assessment and Accreditation of Laboratory Animal Care‐approved temperature (22–24 °C) and humidity (40–60%) controlled colony room. The room was maintained on a 12‐hour light–dark schedule. Pelleted chow and filtered ta...
null
null
null
dx.doi.org/10.17504/protocols.io.iczcax6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.k5ecy3e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><span style="font-weight: 400;">Paraffin embedding and sectioning is a classic technique used in a variety of biological disciplines. Though the fruit fly </span><em><span style="font-weight: 400;">Drosophila melanogaster</span></em><span style="font-weight: 400;"> are one of...
[]
80,849
Assesing Wing Damage Index (WDI) in bats using ImageJ software
1
dx.doi.org/10.17504/protocols.io.j8nlkwq91l5r/v1
https://www.protocols.io/view/assesing-wing-damage-index-wdi-in-bats-using-image-cs7rwhm6
Nia Toshkova, Stanimira Deleva
TITLE: Assesing Wing Damage Index (WDI) in bats using ImageJ software AUTHORS: Nia Toshkova, Stanimira Deleva [DESCRIPTION] Examining the visual health of bats can offer valuable insights into their overall well-being. The condition of wing membranes is essential for bats, supporting water balance and protecting again...
["[Taking bat wing pictures in the field] Place a laminated graph paper on a convenient, flat, and stable surface. The laminated surface will allow you to write down a number code to distinguish between individuals/species and to disinfect it after each individual. Write down a code on the laminated paper containing th...
87,524
A computational pipeline to quantify primary cilia in mouse embryonic fibroblasts with CellProfiler
4
dx.doi.org/10.17504/protocols.io.bp2l6x2j5lqe/v1
https://www.protocols.io/view/a-computational-pipeline-to-quantify-primary-cilia-czqcx5sw
Ebsy Jaimon, Herschel Dhekne, Sreeja Nair, Chloe A Hecht, Suzanne R Pfeffer
TITLE: A computational pipeline to quantify primary cilia in mouse embryonic fibroblasts with CellProfiler AUTHORS: Ebsy Jaimon, Herschel Dhekne, Sreeja Nair, Chloe A Hecht, Suzanne R Pfeffer [DESCRIPTION] We present here an automated CellProfiler (Stirling et al., 2021) software pipeline to quantify the number of pri...
["[Import data into CellProfiler and extract metadata from file names] Select the Images module, drag and drop the maximum intensity projected .TIF files as indicated", "[Group individual channels and create image subsets] Go to Names and types module \nAssign a name to : “Images matching rules” \nProcess as 3D : No\nS...
91,921
Antioxidant and trace metal determinations in human lavage fluids
6
dx.doi.org/10.17504/protocols.io.ewov1qmxygr2/v1
https://www.protocols.io/view/antioxidant-and-trace-metal-determinations-in-huma-c5zry756
Andreas Frølich, i.mudway, anders.blomberg, annelie.behndig
TITLE: Antioxidant and trace metal determinations in human lavage fluids AUTHORS: Andreas Frølich, i.mudway, anders.blomberg, annelie.behndig [DESCRIPTION] Low molecular weight antioxidants (including ascorbate, urate and glutathione), metal transport and chelation proteins (including transferrin and ferritin) and met...
["[Collection and preparation of bronchoalveolar lavage fluid samples for antioxidant analysis:] Bronchoalveolar lavage (BAL) was performed by the instillation and immediate aspiration of 3 consecutive aliquots of 60 mL sodium chloride (NaCl) saline pH 7.3 at 37˚ C into the lingular or middle lobe.\n\nThese recovered a...
29,461
Loop assembly using Labcyte Echo 550
null
dx.doi.org/10.17504/protocols.io.8zvhx66
null
Susana Sauret-Gueto, Anthony West
TITLE: Loop assembly using Labcyte Echo 550 AUTHORS: Susana Sauret-Gueto, Anthony West [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Loop L1 and L2 type IIS assembly protocol, automated and minituarised for assembly of 500nl final volume reactions using the Labcyte Echo 550.</div></div> [STEPS] ?...
["Next create LDV source plate that contain all plasmids needed but the APs.For L1 Loops, have a LDV source plate with the libary of L0 parts.For L2 Loops, have a LDV source plate with the library of L1 plasmids.Use Echo calibration 384LDV_AQ_B2 for these plates.LDV plates working range is~ 3-12 ul (maximum volume is 1...
99,456
Capturing and Processing Slaking Images With a Multi-Well Plate
6
dx.doi.org/10.17504/protocols.io.36wgqjmk5vk5/v3
https://www.protocols.io/view/capturing-and-processing-slaking-images-with-a-mul-ddc822zw
Claire Phillips, Robert E Meadows III, Joaquin Casanova, Bryan Emmett
TITLE: Capturing and Processing Slaking Images With a Multi-Well Plate AUTHORS: Claire Phillips, Robert E Meadows III, Joaquin Casanova, Bryan Emmett [DESCRIPTION] This describes a process to measure soil wet aggregate stability through slaking, or rapid immersion it water. It uses a multi-well plate to process many a...
["[Sample preparation] Establish how you will count the wells (across or down) and create a spreadsheet identifying the sample in each well. \nExample Document:", "[Sample preparation] The well plate we provided a 3D print file for has a notch in the upper left corner to identify the first column and row.", "[Sample pr...
37,563
Use SSH Public Key Authentication
null
null
https://www.protocols.io/view/use-ssh-public-key-authentication-bgw3jxgn
Natapol Pornputtapong
TITLE: Use SSH Public Key Authentication AUTHORS: Natapol Pornputtapong [STEPS] ?. ?.
[]
90,809
GGAssmbler Library Construction
1
dx.doi.org/10.17504/protocols.io.81wgbxqkolpk/v1
https://www.protocols.io/view/ggassmbler-library-construction-c4wzyxf6
Shlomo Yakir Hoch, Ravit Netzer, Karen Hakeny, Sarel J Fleishman
TITLE: GGAssmbler Library Construction AUTHORS: Shlomo Yakir Hoch, Ravit Netzer, Karen Hakeny, Sarel J Fleishman [DESCRIPTION] This protocol describes methods for GGAssembler. [BEFORE_START] Please note that protocols with Q5® High-Fidelity DNA Polymerase may differ from protocols with other polymerases. Conditions r...
["[Amplify constant fragments] Set up the following reaction on ice:", "[Amplify constant fragments] Gently mix the reaction.", "[Amplify constant fragments] Collect all liquid to the bottom of the tube by a quick spin if necessary and overlay the sample with mineral oil if using a PCR machine without a heated lid.", "...
null
null
null
dx.doi.org/10.17504/protocols.io.mpkc5kw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Tongcheng pigs (TC) and Yorkshire (YK) are two pig breeds with distinguished morphologies in muscle. A comprehensive study of porcine microRNAome (miRNAome) in longissimus muscle during 5 developmental stages (40, 55, 63, 70 and 90 dpc (days post coitum)) using Solexa sequenc...
[]
88,186
Chemical ecology in the classroom: chemotaxis assay using C. elegans
4
dx.doi.org/10.17504/protocols.io.e6nvwde9zlmk/v1
https://www.protocols.io/view/chemical-ecology-in-the-classroom-chemotaxis-assay-c2c2yaye
Nicole Bradon, Katherine Fiocca, Maria D. Sallee, Lauren Cote, Daniel A. Shaykevich, Lauren A O'Connell
TITLE: Chemical ecology in the classroom: chemotaxis assay using C. elegans AUTHORS: Nicole Bradon, Katherine Fiocca, Maria D. Sallee, Lauren Cote, Daniel A. Shaykevich, Lauren A O'Connell [DESCRIPTION] There are many natural products in the environment that influence other organisms. How nervous systems sense these c...
["[Obtaining chemical extracts for use as chemotaxis stimuli] Place the collected ants in a freezer for 15 min until immobile.", "[Obtaining chemical extracts for use as chemotaxis stimuli] Wearing gloves, sort ants by species into medium weighing boats.", "[Prepare chemotaxis plates] Label chemotaxis plates.", "[Prepa...
97,853
Preparation, Processing and Preservation of Deceased Donor Kidney Tissue for Multiomic Studies
1
dx.doi.org/10.17504/protocols.io.8epv5r64jg1b/v1
https://www.protocols.io/view/preparation-processing-and-preservation-of-decease-dbs52ng6
Sanjay Jain, Amanda Knoten, Kristy Conlon
TITLE: Preparation, Processing and Preservation of Deceased Donor Kidney Tissue for Multiomic Studies AUTHORS: Sanjay Jain, Amanda Knoten, Kristy Conlon [DESCRIPTION] Multiomic technologies are increasingly being used on human samples and generating multidimensional data. Preanalytical and tissue procurement factors c...
["[Procurement: Procuring Kidney from organ procurement organization (OPO)] Check patient file for the following information:\nConsent to donate organs\nDowntime, if any\nWas kidney on pump?\nBUN/Cr\nKidney biopsy report/any underlying disease\nMedical history for comorbidities\nGeneral good health before the current e...
15,309
Updated QIAGEN PowerSoil protocol
null
dx.doi.org/10.17504/protocols.io.s7mehk6
null
null
TITLE: Updated QIAGEN PowerSoil protocol AUTHORS: [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Adapted from manufacturer instructions for sand extraction for Nanopore sequencing.</div><div class = "text-block">https://www.qiagen.com/au/shop//sample-technologies/dneasy-powersoil-kit/#resources</d...
[]
47,659
SARS-CoV-2 Sequencing on Illumina MiSeq Using ARTIC Protocol: Part 2 - Illumina DNA Prep Protocol
1
null
https://www.protocols.io/view/sars-cov-2-sequencing-on-illumina-miseq-using-arti-bssjnecn
Joel Sevinsky, Arian Nassiri, Heather Blankenship, Erin Young, Kevin Libuit, Kelly Oakeson, Lauren Turner, StaPH-B Consortium
TITLE: SARS-CoV-2 Sequencing on Illumina MiSeq Using ARTIC Protocol: Part 2 - Illumina DNA Prep Protocol AUTHORS: Joel Sevinsky, Arian Nassiri, Heather Blankenship, Erin Young, Kevin Libuit, Kelly Oakeson, Lauren Turner, StaPH-B Consortium [DESCRIPTION] This protocol is an adaption of several circulating protocols on...
["[DNA Prep - Before you begin] Before You Begin\n\nThis protocol is an adaptation of the Illumina DNA Prep kit (below). It is laid out specifically for using amplicons from the ARTIC Tiling PCR Protocol.\n\nThe audience is assumed to be public health laboratorians with access to instruments and reagents used for Puls...
86,877
Immunofluorescence staining for tissue and organoids
4
null
https://www.protocols.io/view/immunofluorescence-staining-for-tissue-and-organoi-cy35xyq6
Gabriela Vallejo Flores, Annika Fendler
TITLE: Immunofluorescence staining for tissue and organoids AUTHORS: Gabriela Vallejo Flores, Annika Fendler [DESCRIPTION] Immunohistochemistry (IHC) is a powerful technique that exploits the specific binding between an antibody and antigen to detect and localize specific antigens in cells and tissue, most commonly ...
["[Blocking] Add blocking buffer A (1% BSA + 2% of host serum (secondary antibody) in TBS 1x, add 150µl for tissue and 50µl for organoids staining, incubate the sample 60 min a room temperature in the humidifying box.", "[Primary antibody] Add the primary antibody to the sample, dilute the antibody in blocking buffer B...
20,088
Scarless introduction of point mutations in mammalian cells with S. pyogenes Cas9
null
dx.doi.org/10.17504/protocols.io.xuyfnxw
null
Malvika Tejura, Stephen Floor
TITLE: Scarless introduction of point mutations in mammalian cells with S. pyogenes Cas9 AUTHORS: Malvika Tejura, Stephen Floor [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol outlines the steps to edit single point mutations into cells in a scarless manner. Uses single-stranded DNA re...
["[Workflow]\nThe workflow for gene editing with the CRISPR/Cas9 system follows these basic steps: Designing appropriate sgRNAs (recruits Cas9 enzyme to DNA). Cas9 bound to the sgRNA is referred to as the \"RNP\" (ribonucleoprotein)Nucleofecting RNPs to test cutting efficiency for each sgRNADesigning appropriate single...
38,921
3: 30mer branch melting temperatures (SABER-FISH)
1
null
https://www.protocols.io/view/3-30mer-branch-melting-temperatures-saber-fish-bh9hj936
Jocelyn Y. Kishi, Sylvain W. Lapan, Brian J Beliveau, Emma R. West, Allen Zhu, Hiroshi M. Sasaki, Sinem Saka, Yu Wang, Constance L Cepko, Peng Yin
TITLE: 3: 30mer branch melting temperatures (SABER-FISH) AUTHORS: Jocelyn Y. Kishi, Sylvain W. Lapan, Brian J Beliveau, Emma R. West, Allen Zhu, Hiroshi M. Sasaki, Sinem Saka, Yu Wang, Constance L Cepko, Peng Yin [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes the design of 3...
["It's recommended to use a temperature at least 1 degree lower than the lowest melting temperature of all branch sequences you plan to use (see plot of melting temperatures Fig. S2).You can find these melting temperature curves reported for each sequence, as well as those computed for 20mer imagers, 42mer barcode sequ...
98,441
Urine Lipoarabinomannan as a Biomarker for Mycobacterium tuberculosis and non-tuberculous mycobacterial infections
0
dx.doi.org/10.17504/protocols.io.3byl497o8go5/v1
https://www.protocols.io/view/urine-lipoarabinomannan-as-a-biomarker-for-mycobac-dcdh2s36
Jordan Manzer, Ibrahim Aboellail, Jerry Nick, Delphi Chatterjee, Anita G Amin
TITLE: Urine Lipoarabinomannan as a Biomarker for Mycobacterium tuberculosis and non-tuberculous mycobacterial infections AUTHORS: Jordan Manzer, Ibrahim Aboellail, Jerry Nick, Delphi Chatterjee, Anita G Amin [DESCRIPTION] Lipoarabinomannan (LAM) is a unique cell wall component containing a fatty acid region with a la...
["[Urinary LAM Tuberculostearic acid Analysis] Hexane Extraction (Removes exogenous proteins and lipids from urine)", "[Urinary LAM Tuberculostearic acid Analysis] Using 1 mL of urine in a KIMAX 13 x 100mm glass tube with teflon screw cap, 1 mL of Hexane is added.", "[Urinary LAM Tuberculostearic acid Analysis] Vortex ...
95,955
Viral Enumeration of Water Samples Using Wet Mount Epifluorescence Microscopy
0
dx.doi.org/10.17504/protocols.io.q26g7po68gwz/v2
https://www.protocols.io/view/viral-enumeration-of-water-samples-using-wet-mount-c9xtz7nn
Madeline Bellanger, Pieter Visscher, Richard Allen White III
TITLE: Viral Enumeration of Water Samples Using Wet Mount Epifluorescence Microscopy AUTHORS: Madeline Bellanger, Pieter Visscher, Richard Allen White III [DESCRIPTION] Epifluorescence microscopy (EFM) has been the gold standard method for environmental viral enumeration for over 25 years. Currently, standard EFM meth...
["[Filtration and Cleaning] Record the starting volume of water, this will be used for calculations later.", "[Filtration and Cleaning] Filter water through a glass fiber pre filter (Whatman Grade 1 qualitative filter paper).", "[Filtration and Cleaning] Collect filtrate and filter through a 0.65 μm PVDF filter (Durapo...
85,959
Mitochondria purification
4
dx.doi.org/10.17504/protocols.io.n92ldmppnl5b/v1
https://www.protocols.io/view/mitochondria-purification-cx7fxrjn
wusj, schekman
TITLE: Mitochondria purification AUTHORS: wusj, schekman [DESCRIPTION] This protocol describes how to purify mitochondria from cell culture [STEPS] SECTION: Mitochondria purification 1. HEK293T cells were trypsinized (0.05% Trypsin 5 min ) and collected by centrifugation (500g 5 min). SECTION: Mitochondria purifica...
["[Mitochondria purification] HEK293T cells were trypsinized (0.05% Trypsin 5 min ) and collected by centrifugation (500g 5 min).", "[Mitochondria purification] Cells were washed twice with NKM buffer (1 mM Tris HCl, pH7.3, 0.13 M NaCl, 5 mM KCl, and 7.5 mM MgCl2), and resuspended in six packed cell volumes of homogeni...
103,664
LTEE Media Recipes 
1
dx.doi.org/10.17504/protocols.io.81wgbyr31vpk/v4
https://www.protocols.io/view/ltee-media-recipes-dhgq33vw
Jesus E Chavarria-Palma, Zachary D Blount, Jeffrey E Barrick
TITLE: LTEE Media Recipes  AUTHORS: Jesus E Chavarria-Palma, Zachary D Blount, Jeffrey E Barrick [DESCRIPTION] This protocol describes recipes to prepare growth media and reagents used in the E. coli long-term evolution experiment (LTEE). Section 1: DM-glucose, Davis-Mingioli liquid medium supplemented with glucose ...
["[DM-glucose: Davis-Mingioli liquid medium supplemented with glucose] To prepare 1 L of DM, follow these steps.", "[DM-glucose: Davis-Mingioli liquid medium supplemented with glucose] Weigh dry components:\na. 5.34 g of or 7 gof \nb. 2 g of \nc. 1 g of \nd. 0.5 g of", "[DM-glucose: Davis-Mingioli liquid medi...
28,773
Copy of Fabrication of Microneedle Patches
null
dx.doi.org/10.17504/protocols.io.8cdhss6
null
Stephania Konstantinidi, Hana Samet
TITLE: Copy of Fabrication of Microneedle Patches AUTHORS: Stephania Konstantinidi, Hana Samet [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The goal is to create a microneedle. patch, which, by applying it on a leaf, will extract amplification-ready DNA.</div><div class = "text-block">All MN patc...
["[Mold preparation]\nClean the polydimethylsiloxane molds.", "[Mold preparation]\nHeat deionised water to 100 degrees Celsius using a becher.\n100 °C", "[Mold preparation]\nPut the mold(s) into the water and stir softly.", "[Mold preparation]\nRemove the molds and let them dry in a chemical hood for 1-2 hours.", "[M...
108,296
Mass Spectrometry Identification of Samples Separated by Liquid Chromatography
0
dx.doi.org/10.17504/protocols.io.bp2l625kdgqe/v1
https://www.protocols.io/view/mass-spectrometry-identification-of-samples-separa-dmzg473w
Caroline Green
TITLE: Mass Spectrometry Identification of Samples Separated by Liquid Chromatography AUTHORS: Caroline Green [DESCRIPTION] As one of the most powerful separation and analysis technologies in the field of chemistry, liquid chromatography (LC) has developed rapidly since the 1970s. Both in terms of fundamental chromato...
["[Guide for Mass Spectrometry Identification of Samples Separated by Liquid Chromatography] Main Instruments and Equipment", "[Guide for Mass Spectrometry Identification of Samples Separated by Liquid Chromatography] Main reagents*1", "[Guide for Mass Spectrometry Identification of Samples Separated by Liquid Chromato...
67,433
Keto Complete Australia {AU/UK} Pills Reviews!
3
dx.doi.org/10.17504/protocols.io.81wgb631olpk/v1
https://www.protocols.io/view/keto-complete-australia-au-uk-pills-reviews-cd4hs8t6
Keto Complete Australia
TITLE: Keto Complete Australia {AU/UK} Pills Reviews! AUTHORS: Keto Complete Australia [DESCRIPTION] Keto Complete Australia [STEPS]
[]
66,634
Microscopy-based bead protein-protein interaction assay
4
dx.doi.org/10.17504/protocols.io.6qpvr6nwpvmk/v1
https://www.protocols.io/view/microscopy-based-bead-protein-protein-interaction-cdbis2ke
Justyna Sawa-Makarska
TITLE: Microscopy-based bead protein-protein interaction assay AUTHORS: Justyna Sawa-Makarska [DESCRIPTION] This protocol describes how to perform microscopy-based bead protein-protein interaction assay with GST-labelled proteins as baits and fluorescently labelled proteins as preys. The protocol requires to have puri...
["[Interaction assay] Pipette the preys onto 384-well glass-bottom microplate (Greiner Bio-One). The minimal volume to cover the bottom of a well is 20 μl. The final concentration of the prey may vary depending on the strength of the interaction, usually between 0.1-1 μM. The number of proteins in the well can be adjus...
null
null
null
dx.doi.org/10.17504/protocols.io.dpt5nm
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] A generally more efficient way of isolating lytic phages from marine environments is by the use of enrichment cultures. In this approach, the prefiltered water sample that is to be screened for phages against a given target bacterium, is enriched with a bacterial growth medium a...
[]
94,458
Tissue sampling from museum specimens of mitten crabs
4
dx.doi.org/10.17504/protocols.io.6qpvr3n1zvmk/v1
https://www.protocols.io/view/tissue-sampling-from-museum-specimens-of-mitten-cr-c8g2ztye
Christine Ewers
TITLE: Tissue sampling from museum specimens of mitten crabs AUTHORS: Christine Ewers [DESCRIPTION] Simple protocol for sampling tissues from mitten crabs housed in natural history collections. Mitten crabs are somewhat unique because they have hair on their chelae from which DNA can be extracted. However, the protoco...
["Clean all surfaces with DNAaway", "Flame forceps and scalpel", "Open collection jar", "Take out one specimen", "Place in clean tray with scale (e.g. ruler). Take first picture of the collection metadata (the jar label) and then picture of the crab. This way you can trace back which crab you photographed", "Chose the ...
69,934
Stereology-mediated cell count using StereoInvestigator
4
dx.doi.org/10.17504/protocols.io.6qpvr4ekbgmk/v1
https://www.protocols.io/view/stereology-mediated-cell-count-using-stereoinvesti-cgintude
miquel.vila
TITLE: Stereology-mediated cell count using StereoInvestigator AUTHORS: miquel.vila [DESCRIPTION] Protocol for cell counting using StereoInvestigator software [STEPS] 1. Set slide properly on the microscope and set new reference point 2. Click on Probes → Optical Fractionator Workflow → Start a new subject → 3. Ente...
["Set slide properly on the microscope and set new reference point", "Click on Probes → Optical Fractionator Workflow → Start a new subject →", "Enter cut thickness (5 µm (cut with microtome, 20 µm or 30 µm (cut with cryostat))", "Enter interval according to the thickness: 17 if 5 µm, 6 if 20 µm or 4 if µm)", "In Sele...
86,180
iNDI Papain Dissociation protocol for iNeurons Version 1
4
dx.doi.org/10.17504/protocols.io.kxygx392dg8j/v1
https://www.protocols.io/view/indi-papain-dissociation-protocol-for-ineurons-ver-cyecxtaw
Erika Lara Flores, Joel Reyes, Mark Cookson, Michael Ward
TITLE: iNDI Papain Dissociation protocol for iNeurons Version 1 AUTHORS: Erika Lara Flores, Joel Reyes, Mark Cookson, Michael Ward [DESCRIPTION] Proteolytic enzymes are widely used in cell dissociation and papain has proved less damaging with some tissues and neuronal cultures and more effective than other proteases....
["[Preparation of Reagents] Equilibrate EBSS/phenol red with 95%O2:5%CO2 in incubator for several hours of overnight.", "[Preparation of Reagents] Reconstitute the Ovomucoid mixture (Papain inhibitor-PI) by adding 32 mL of EBSS and allow contents to dissolve. This will yield solution at an effective concentration of 10...
75,168
Measles whole-genome sequencing
4
null
https://www.protocols.io/view/measles-whole-genome-sequencing-cmm8u49w
My VT Phan, Matthew Cotten
TITLE: Measles whole-genome sequencing AUTHORS: My VT Phan, Matthew Cotten [DESCRIPTION] This is an optimised protocol for PCR amplification of the Measles virus (MV) genome using specific primers designed for B3 genotype. The method generates cDNA amplicons suitable for whole genome sequencing using the MinION platfo...
["[PCR amplification of measles virus genome] cDNA synthesis", "[PCR amplification of measles virus genome] For each FPM reaction, add the following:\n4 µL \n1 µL \n1 µL \n1 µL \n1 µL \n\nThen incubate the reactions using the following conditions:\n42 °C for 50 min \n70 °C for 10 min", "[PCR amplification of measles v...
25,838
Spectrophotometric assay of the mitochondrial F1F0 ATP synthase
null
dx.doi.org/10.17504/protocols.io.5gng3ve
null
Anne LOMBES, Francis Haraux
TITLE: Spectrophotometric assay of the mitochondrial F1F0 ATP synthase AUTHORS: Anne LOMBES, Francis Haraux [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "vertical-align:sub;">F</span><span style = "vertical-align:sub;">1</span><span style = "vertical-align:sub;vertical-align:sub;">F...
["Reagents preparation 1- Homogenization buffer (kept at 0-4°C for few weeks or at -20°C for longer period of time)= 225 mM mannitol, 75 mM sucrose, 0.1 mM EDTA, and 10 mM Tris HCl pH 7.2Anti-protease cocktail at the working concentration given by the manufacturer should be added before use2- Assay buffer (kept at 0-4°...
40,896
ELISA for quantification of interferon gamma (IFN-γ) in human serum or plasma.
6
dx.doi.org/10.17504/protocols.io.bj68krhw
https://www.protocols.io/view/elisa-for-quantification-of-interferon-gamma-ifn-bj68krhw
Angel Justiz-Vaillant, Belkis Ferrer-Cosme
TITLE: ELISA for quantification of interferon gamma (IFN-γ) in human serum or plasma. AUTHORS: Angel Justiz-Vaillant, Belkis Ferrer-Cosme [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">IFN-γ is a key player in driving cellular immunity. It is capable of orchestrating numerous and vital prot...
["An anti-human IFN-γ 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 IFN-γ present in the serum or plasma binds to antibodies adsorbed into the microwells.", "The microplate is blocked with 3% non-fat milk-P...
46,787
Artificial sea water 
4
dx.doi.org/10.17504/protocols.io.brxbm7in
https://www.protocols.io/view/artificial-sea-water-brxbm7in
Simon Blanchoud
TITLE: Artificial sea water  AUTHORS: Simon Blanchoud [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Three alternative solutions for artificial seawater (ASW) have been tested successfully on our </span><span style = "font-style:italic;">Botrylloides</span><span> colonies. For routine work, ...
["[2X CSS]\nsea salt in H2O.\n70 g\n1 L", "[2X CSS]\nMix to dissolve.", "[2X CSS]\nFilter at 10 µm", "[10x CSHP]\nTo prepare 10X CSHP take NaCl262.9g4.5MKCl7.4g100mMCaCl29.9g90mMMgCl2 6H2O60.9g300mMMgSO4 7H2O39.4g160mMH2O1000ml\nNaCl262.9g4.5MKCl7.4g100mMCaCl29.9g90mMMgCl2 6H2O60.9g300mMMgSO4 7H2O39.4g160mMH2O1000ml",...
42,595
NOAA-CalCOFI Ocean Genomics (NCOG) Sample Collection
1
dx.doi.org/10.17504/protocols.io.bmubk6sn
https://www.protocols.io/view/noaa-calcofi-ocean-genomics-ncog-sample-collection-bmubk6sn
Ariel Rabines, Rob Lampe, Lisa Zeigler Allen, Andrew Allen
TITLE: NOAA-CalCOFI Ocean Genomics (NCOG) Sample Collection AUTHORS: Ariel Rabines, Rob Lampe, Lisa Zeigler Allen, Andrew Allen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol describes our sampling strategy and techniques for the </span><a href="https://calcofi.org/LRbLa/field-w...
["[Station sampling and depths]\nFor station positions and cruise patterns, refer to the CalCOFI website. Stations:Prodo stations refers to stations sampled around the same time of day (late morning) when primary productivity is measured. Cardinal stations refers to stations at a certain location to be sampled no matte...
null
null
null
dx.doi.org/10.17504/protocols.io.j9qcr5w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Diet-induced obesity through administration of a western-type diet high in fat and sugar to mice can result in a higher weight of adipose and liver tissues. The increased adipose tissue weight can be due to hypertrophy (bigger cells) or hyperplasia (more cells) of adipocytes,...
[]
89,921
WesternBlot Paramecium bursaria (Pb)
1
dx.doi.org/10.17504/protocols.io.ewov1q58kgr2/v1
https://www.protocols.io/view/westernblot-paramecium-bursaria-pb-c329yqh6
Estelle Kilias
TITLE: WesternBlot Paramecium bursaria (Pb) AUTHORS: Estelle Kilias [DESCRIPTION] Western Blot protocol for Pb including all steps from the cell harvest to the final ECL stain. Culture density is an important factor that requires checking prior starting. [STEPS] SECTION: Protein extraction 1. To concentrate the cultu...
["[Protein extraction] To concentrate the culture, strain 100 ml of a dense Paramecium culture through a 40 micrometer strainer and split the volume over two 50 ml falcon tubes", "[Protein extraction] Centrifuge the strained culture for 10 min at 800xg", "[Protein extraction] Remove the supernatant from each falcon tub...
71,457
Immunostaining of H&amp;E Stained Paraffin Sections of Fly Heads
4
dx.doi.org/10.17504/protocols.io.81wgby61nvpk/v1
https://www.protocols.io/view/immunostaining-of-h-amp-e-stained-paraffin-section-chz9t796
Mel Feany
TITLE: Immunostaining of H&amp;E Stained Paraffin Sections of Fly Heads AUTHORS: Mel Feany [DESCRIPTION] This protocol describes how to perform immunostaining on H&E stained paraffin sections of fly heads. [STEPS] 1. Embed the fly heads in paraffin and stain with H&E for neuronal counting, per the following protocol:...
["Embed the fly heads in paraffin and stain with H&E for neuronal counting, per the following protocol:\ndx.doi.org/10.17504/protocols.io.4r3l24on4g1y/v1", "Microwave slides in 10 millimolar (mM) sodium citrate for 15 min. \nCool 20 min .\n\nStock: 100 millimolar (mM) sodium citrate, pH 6.0\nUse at least 1 L of citra...
101,120
Purification of Lambda Protein Phosphatase
0
dx.doi.org/10.17504/protocols.io.kqdg322bqv25/v1
https://www.protocols.io/view/purification-of-lambda-protein-phosphatase-dey83fzw
Elias Adriaenssens
TITLE: Purification of Lambda Protein Phosphatase AUTHORS: Elias Adriaenssens [DESCRIPTION] This protocol details the purification of Lambda protein phosphatase. [STEPS] SECTION: Purification procedure 1. To purify Lambda protein phosphatase (λ PPase), fuse the protein phosphatase to a N-terminal 6xHis-tag through cl...
["[Purification procedure] To purify Lambda protein phosphatase (λ PPase), fuse the protein phosphatase to a N-terminal 6xHis-tag through cloning into a pET-DUET1 vector (available from Addgene).", "[Purification procedure] After the transformation of the pET-DUET1 vector encoding 6xHis-TEV-λ PPase in E. coli Rosetta p...
79,337
Adhesion assay
1
dx.doi.org/10.17504/protocols.io.dm6gpj111gzp/v1
https://www.protocols.click/view/adhesion-assay-crqhv5t6
Philippa R Kennedy, Quinlan Kile
TITLE: Adhesion assay AUTHORS: Philippa R Kennedy, Quinlan Kile [DESCRIPTION] Adapted from Natural Killer Cell Protocols 2009 p89-96 Enriched effector cells and tumor targets are labelled with different fluorescence. They are allowed to interact over a time-course and samples are taken and fixed at regular intervals....
["[Set aside cells for flow cytometer setup] The following controls will be required to setup the gates on a flow cytometer:\n1. Unlabeled cells for an unstained control\n2. Single color control (effector) e.g. CellTrace Violet labeled NK cells.\n3. Single color control (target) e.g. K562-Nuclight Red.", "[Prepare the ...
58,605
Utilization Patterns and Trends in The Use of Medications for Asthma Control in a Cohort of Colombian Patients, 2017-2019
1
dx.doi.org/10.17504/protocols.io.b5gmq3u6
https://www.protocols.io/view/utilization-patterns-and-trends-in-the-use-of-medi-b5gmq3u6
Jorge Machado Alba
TITLE: Utilization Patterns and Trends in The Use of Medications for Asthma Control in a Cohort of Colombian Patients, 2017-2019 AUTHORS: Jorge Machado Alba [DESCRIPTION] Background:Asthma affects approximately 358 million people worldwide. Objective:To determine the trend in the use of medications used to trea...
[]
45,989
Understanding the impact of sole soled footwear on toddler spatial, temporal and kinetic gait parameters.
1
dx.doi.org/10.17504/protocols.io.bq6dmza6
https://www.protocols.io/view/understanding-the-impact-of-sole-soled-footwear-on-bq6dmza6
Cylie Williams, Jessica Kolic, Wen Wu, Kade Paterson
TITLE: Understanding the impact of sole soled footwear on toddler spatial, temporal and kinetic gait parameters. AUTHORS: Cylie Williams, Jessica Kolic, Wen Wu, Kade Paterson [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A quasi-experimental pre-post study design will be used to assess whether sp...
["Aim: To assess whether spatiotemporal parameters of gait, and in-shoe foot and lower limb kinematics differ when walking barefoot and in soft-soled footwear in newly walking toddlers. Hypothesis: Soft soled footwear would result in similar differences in gait variables as those observed in older children (ages 2-4)."...
62,104
Rehabilitation interventions for impaired handwriting in people with Parkinson’s disease: a protocol for a scoping review
1
dx.doi.org/10.17504/protocols.io.5jyl891z8v2w/v1
https://www.protocols.io/view/rehabilitation-interventions-for-impaired-handwrit-b8vyrw7w
Andrea Gardoni, Elisabetta Sarasso, Federica Agosta, Massimo Filippi, Davide Corbetta
TITLE: Rehabilitation interventions for impaired handwriting in people with Parkinson’s disease: a protocol for a scoping review AUTHORS: Andrea Gardoni, Elisabetta Sarasso, Federica Agosta, Massimo Filippi, Davide Corbetta [DESCRIPTION] Handwriting abnormalities in people with Parkinson’s disease include several dyn...
["[Search strategy] The search strategy was runned in April 2022", "[Study records management, selection and data collection]", "[Risk of bias assessment] Requested time will depend on how many studies will be included", "[Data synthesis] Requested time will depend on how many studies will be included"]
50,537
Flow Cytometry ECS Surface Antigens
4
dx.doi.org/10.17504/protocols.io.bvkhn4t6
https://www.protocols.io/view/flow-cytometry-ecs-surface-antigens-bvkhn4t6
Michael Betts, Gregory Golden
TITLE: Flow Cytometry ECS Surface Antigens AUTHORS: Michael Betts, Gregory Golden [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">High-parameter flow cytometry enables identification and characterization of a wide range of cell populations within a biological sample. Analysis of cell-surface epitope...
["[Procedure]\nThawing and Restinga. Pre-warm R10 media in a water bath. b. Thaw samples in-vial using a water bath.c. Add thawed cells to of R10, then spin cells at 500 xg for 5 min.d. Resuspend cell pellet in of R10 and count cells. e. Rest cells at least 3 hours (up to overnight) at 2x106 cells/mL in R10 medium ...
105,953
Osmia lignaria laboratory rearing protocol
1
dx.doi.org/10.17504/protocols.io.eq2lyj4qplx9/v2
https://www.protocols.io/view/osmia-lignaria-laboratory-rearing-protocol-djp94mr6
Mary-Kate F. Williams, Natalie K. Boyle, Robert N. Schaeffer, Diana L Cox-Foster
TITLE: Osmia lignaria laboratory rearing protocol AUTHORS: Mary-Kate F. Williams, Natalie K. Boyle, Robert N. Schaeffer, Diana L Cox-Foster [DESCRIPTION] Our protocol was designed to rear Osmia lignaria Say (Hymenoptera: Megachilidae) from immature stages to adult emergence following their natural phenology in norther...
["[Propagation and acquiring solitary bee nests (Fig. 1)] Deploy corrugated plastic nesting boxes (Artz et al. 2013, 2014) into the orchard before bloom. Place nest blocks with straw inserts or bundles of cardboard tubes with straw inserts into nest boxes.", "[Propagation and acquiring solitary bee nests (Fig. 1)] Once...
49,490
Fixation of HeLa-M cells expressing Halo and SNAP fusion proteins conjugated to ligands
4
dx.doi.org/10.17504/protocols.io.bujsnune
https://www.protocols.io/view/fixation-of-hela-m-cells-expressing-halo-and-snap-bujsnune
OLIVIA HARDING, Erika L. F. Holzbaur
TITLE: Fixation of HeLa-M cells expressing Halo and SNAP fusion proteins conjugated to ligands AUTHORS: OLIVIA HARDING, Erika L. F. Holzbaur [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Mitophagy is a tightly regulated mechanism in which components are sequentially recruited to a damaged mitocho...
["[Before start]\nThe start point for this protocol is after cells grown on a glass coverslip in a 12- well plate have been transfected with YFP-Parkin, Halo-OPTN, and SNAP-TBK1 for - .", "[Before start]\nPrepare in PBS. Keep frozen at .\n[PFA]\n-20 °C\nPrepare fresh for day-of fixation or thaw directly before use.\n...
17,062
ChroDrip - ProteinA
null
dx.doi.org/10.17504/protocols.io.uweexbe
null
David Frommholz, Alexandra Ehl, Nadine Stefanczyk
TITLE: ChroDrip - ProteinA AUTHORS: David Frommholz, Alexandra Ehl, Nadine Stefanczyk [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Purification Guide for the Isolation of Antibodies with ChroDrip Columns by DALEX Biotech.</span></div><div class = "text-block">Eas...
["[Load and Wash]\nAdd your sample to the top of the column and let it flow through by gravity.\nFor optimal binding and purity, the pH of the sample should be 7.5 - 8.5 and should contain 150 - 300 mM NaCl. An easy way to achieve this is by adding 1/11 volume 0.5 M Tris, 2 M NaCl (pH 8.0) to your sample.Removal of par...
88,357
Protocol for abscisic acid (ABA) extraction from plant seeds
4
dx.doi.org/10.17504/protocols.io.81wgbxnnqlpk/v1
https://www.protocols.io/view/protocol-for-abscisic-acid-aba-extraction-from-pla-c2idyca6
Jian You Wang, Lamis Berqdar, Salim Al-Babili
TITLE: Protocol for abscisic acid (ABA) extraction from plant seeds AUTHORS: Jian You Wang, Lamis Berqdar, Salim Al-Babili [DESCRIPTION] The plant hormone abscisic acid (ABA) regulates seed dormancy and germination. Here, we present a protocol for ABA extraction from plant seeds. We describe necessary steps required f...
["[Sample preparation] Grind 7-8 seeds in a Safe-Lock 2.0 mL Eppendorf tube with 3-4 beads for 1 min – frequency 25-26 Hz.", "[Sample preparation] Weight 100 mg fine powder of ground seeds.", "[Sample preparation] Add 1 mL of Standard solution in each tube.", "[Sample preparation] Sonicate the samples for 15 min.", "[S...
40,368
Inhibition immunoassay of the HIVgp120_anti-HIVgp120 (Ab-1) reaction by anti-anti-idiotypic-HIVgp120 antibody (Ab-3).
4
dx.doi.org/10.17504/protocols.io.bjnqkmdw
https://www.protocols.io/view/inhibition-immunoassay-of-the-hivgp120-anti-hivgp1-bjnqkmdw
Angel Justiz-Vaillant
TITLE: Inhibition immunoassay of the HIVgp120_anti-HIVgp120 (Ab-1) reaction by anti-anti-idiotypic-HIVgp120 antibody (Ab-3). AUTHORS: Angel Justiz-Vaillant [STEPS] ?. Ninety six well polystyrene microplate (U-shaped bottom, Sigma-Aldrich Co, St Louis Missouri) is coated with 50 µl/well of 1 ng/µl solution of gp-120...
["Ninety six well polystyrene microplate (U-shaped bottom, Sigma-Aldrich Co, St Louis Missouri) is coated with 50 µl/well of 1 ng/µl solution of gp-120 peptide, fragment 254-274 (Sigma) in carbonate-bicarbonate buffer pH 9.6 (Sigma) for 4 hours at 37°C.", "The microplate is then washed 4 times with PBS Tween-20.", "The...
28,992
Crude extraction of Xylella fastidiosa from infected leaves
null
dx.doi.org/10.17504/protocols.io.8i8huhw
null
Niels Appelman
TITLE: Crude extraction of Xylella fastidiosa from infected leaves AUTHORS: Niels Appelman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol can be used to (crudely) extract Xylella fastidiosa from infected leaves.</div></div> [STEPS] ?. Pick leaves from plants infected with X. fastidio...
["Pick leaves from plants infected with X. fastidiosa", "With a scalpel, remove the first 2 cm of the central nerve of the plant was removed.Note: X. fastidiosa is located in the xylem in this nerve. Including mesophyll will reduce the yield.", "Crush the central nerve and extract bacteria with 50 ml of 0.01 M potassiu...
null
null
null
dx.doi.org/10.17504/protocols.io.shteb6n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The importance of fungal species differentiation is critical for plant pathology research and disease control purposes. Traditionally, fungal species have been identified by a range of cultural and morphological characteristics, such as conidial morphology, presence or absenc...
[]
38,918
SABER-FISH – Signal amplification for multiplexed fluorescence in situ hybridization assays
2
null
https://www.protocols.io/view/saber-fish-signal-amplification-for-multiplexed-fl-bh9ej93e
Jocelyn Y. Kishi, Sylvain W. Lapan, Brian J Beliveau, Emma R. West, Allen Zhu, Hiroshi M. Sasaki, Sinem Saka, Yu Wang, Constance L Cepko, Peng Yin
TITLE: SABER-FISH – Signal amplification for multiplexed fluorescence in situ hybridization assays AUTHORS: Jocelyn Y. Kishi, Sylvain W. Lapan, Brian J Beliveau, Emma R. West, Allen Zhu, Hiroshi M. Sasaki, Sinem Saka, Yu Wang, Constance L Cepko, Peng Yin [DESCRIPTION] <div class = "text-blocks"><div class = "text-bloc...
[]
50,401
RT–PCR protocol for the detection ORF1 11288–11296 deletion (NSP6 106-108del) in SARS-CoV-2 genome
4
dx.doi.org/10.17504/protocols.io.bvf9n3r6
https://www.protocols.io/view/rt-pcr-protocol-for-the-detection-orf1-11288-11296-bvf9n3r6
Nikita Yolshin , Artem Fadeev, Andrey Komissarov
TITLE: RT–PCR protocol for the detection ORF1 11288–11296 deletion (NSP6 106-108del) in SARS-CoV-2 genome AUTHORS: Nikita Yolshin , Artem Fadeev, Andrey Komissarov [DESCRIPTION] ORF1 deletion is observed in some new SARS-CoV-2 lineages and is lineage defining for B.1.1.7 and P.1. Biological effect of this mutation ...
["Order oligonucleotides with following sequences:\n\n \n Name Sequence (5’ – 3’) ORF1-del-F GGTTGGATATGGTTGATACTAGTTTGAAG ORF1-del-R TGTCAAGACATTCATAAGTGTCCACA ORF1-del-P Cy5.5-ACTGTGTTATGTATGCATCAGCTGTAGTGTTACTAATCC-BHQ3 \n \n 2X RT-PCR Buffer 12.5 μl ...
35,927
Quant-iT™ RiboGreen™ RNA Quantification
null
dx.doi.org/10.17504/protocols.io.bfbxjipn
https://www.protocols.io/view/quant-it-ribogreen-rna-quantification-bfbxjipn
Roey Angel, Eva Petrova
TITLE: Quant-iT™ RiboGreen™ RNA 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 RNA using </span><a href="https://www.thermofisher.com/order/catalog/product/R11490#/R11490" style = "tex...
["[Prepare the reaction]\nTake out all reagents from the fridge and bring them to room temperature.Take out the RNA samples from the freezer. RNA samples should be slowly thawed on ice.\nQuant-iT™ RiboGreen® RNA reagent is dissolved in dimethylsulfoxide (DMSO), which freezes below 19 °C. The reagent must be completely ...
40,359
Protein quantification protocol optimized for zebrafish brain tissue (Bradford method)
6
dx.doi.org/10.17504/protocols.io.bjnfkmbn
https://www.protocols.io/view/protein-quantification-protocol-optimized-for-zebr-bjnfkmbn
Adrieli Sachett, Matheus Gallas-Lopes, Greicy M M Conterato, Radharani , Ana Herrmann, Angelo Piato
TITLE: Protein quantification protocol optimized for zebrafish brain tissue (Bradford method) AUTHORS: Adrieli Sachett, Matheus Gallas-Lopes, Greicy M M Conterato, Radharani , Ana Herrmann, Angelo Piato [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Zebrafish are incresingly used as a model animal...
["[Preparing the reagents]\nThe first step is to prepare the reagents to be used in protein quantification;", "[Preparing the reagents]\nBradford reagent: Prepare and use this reagent under dim or no light, making sure all glassware used is covered in aluminum foil to avoid photodegradation of the reagent. Prepare and ...
104,065
Automated Bar-Seq Library Preparation and Pooling
0
dx.doi.org/10.17504/protocols.io.3byl49qdjgo5/v2
https://www.protocols.io/view/automated-bar-seq-library-preparation-and-pooling-dhu936z6
David Ross, Nina Alperovich
TITLE: Automated Bar-Seq Library Preparation and Pooling AUTHORS: David Ross, Nina Alperovich [DESCRIPTION] Protocol for automated Bar-Seq Library preparation This protocol prepares 96 DNA samples, representing 24 samples from 4 different timepoints, for multiplexed Illumina sequencing. The process starts with two r...
["[Transfer Plasmid DNA to PCR Plate and Dilute with DI water] Pre-heat the on deck thermocycler (ODTC) for 1st PCR step", "[Transfer Plasmid DNA to PCR Plate and Dilute with DI water] Remove lids from PCR-Plate 1, Sample-Plate, and Reagent Plate", "[Transfer Plasmid DNA to PCR Plate and Dilute with DI water] Transfer ...
55,913
Study Protocol Document for PONE-D-21-18427R2 (Prevalence and Predictors of GMD among PLWH on cART)
3
dx.doi.org/10.17504/protocols.io.b2uhqet6
https://www.protocols.io/view/study-protocol-document-for-pone-d-21-18427r2-prev-b2uhqet6
Wondmagegn T. Tadesse, Wondwossen Amonge, Eleni Akllilu, Ephrem Engidawork
TITLE: Study Protocol Document for PONE-D-21-18427R2 (Prevalence and Predictors of GMD among PLWH on cART) AUTHORS: Wondmagegn T. Tadesse, Wondwossen Amonge, Eleni Akllilu, Ephrem Engidawork [DESCRIPTION] This is a document that states about sample size calculation, study participant recruitment, interview and me...
[]
84,890
FACS for CD45+ immune cells
4
dx.doi.org/10.17504/protocols.io.36wgq3465lk5/v1
https://www.protocols.click/view/facs-for-cd45-immune-cells-cw52xg8e
Connor Monahan
TITLE: FACS for CD45+ immune cells AUTHORS: Connor Monahan [DESCRIPTION] This protocol details FACS of intestinal immune cells (CD45+). [STEPS] SECTION: Procedure 1. Resuspended cells isolated from the small intestine with 400 µL Fc block (BD Biosciences, Cat# 553142; Franklin Lakes, NJ), 1:200 in FACS buffer. SECTIO...
["[Procedure] Resuspended cells isolated from the small intestine with 400 µL Fc block (BD Biosciences,\nCat# 553142; Franklin Lakes, NJ), 1:200 in FACS buffer.", "[Procedure] Incubate for 20 min on ice.", "[Procedure] Wash cells in 500 µL FACS buffer, pellet at 1500 rpm, 2 min, then resuspend in 600 µL of FACS buffer ...
91,169
ReViBE: protocol for Refit Visualisation of lithic reduction sequences using the Blender Engine
1
dx.doi.org/10.17504/protocols.io.ewov1qxqkgr2/v1
https://www.protocols.io/view/revibe-protocol-for-refit-visualisation-of-lithic-c499yz96
Javier Sánchez-Martínez, Katia Calmet, Jorge Martínez-Moreno, Xavier Roda Gilabert
TITLE: ReViBE: protocol for Refit Visualisation of lithic reduction sequences using the Blender Engine AUTHORS: Javier Sánchez-Martínez, Katia Calmet, Jorge Martínez-Moreno, Xavier Roda Gilabert [DESCRIPTION] Here, we introduce ReViBE, a step-by-step protocol for visualising lithic refits through the utilisation of im...
["[Part 1 - Initial remarks and set up preparation] Place the camera on a tripod (ideally with a zoom lens with a focal length between 35mm and 80mm). Do not use an autofocus lens.", "[Part 1 - Initial remarks and set up preparation] With 3 light sources, create diffused lighting from both sides and above. Try to avoid...
69,485
Integra Magbead DNA and RNA Extraction for isolated colonies 
1
dx.doi.org/10.17504/protocols.io.dm6gpjeqjgzp/v1
https://www.protocols.io/view/integra-magbead-dna-and-rna-extraction-for-isolate-cf4mtqu6
Sophana Chea, Sreyngim Lay, Mengheng Oum, Gechlang Tang, Cheata Hou, Manu Vanaerschot, Christina Yek, Cristina Tato, Jessica Manning, Vida Ahyong
TITLE: Integra Magbead DNA and RNA Extraction for isolated colonies  AUTHORS: Sophana Chea, Sreyngim Lay, Mengheng Oum, Gechlang Tang, Cheata Hou, Manu Vanaerschot, Christina Yek, Cristina Tato, Jessica Manning, Vida Ahyong [DESCRIPTION] This protocol is the process to extract DNA and RNA from isolated colonies. The e...
["[Buffer Preparation] 1. Add 20 mL isopropanol to the MagBead DNA/RNA Wash 1 concentrate.\n2. Add30 mL isopropanol to the MagBead DNA/RNA Wash 2 concentrate.\n3. Reconstitute lyophilized Proteinase K at 20 mg/mL with Proteinase K Storage Buffer and mix by vortexing. Use immediately or store at -20 °C .\n4. Reconstitut...
48,817
Woodchip Ashing
1
null
https://www.protocols.io/view/woodchip-ashing-btwrnpd6
Feyereisen
TITLE: Woodchip Ashing AUTHORS: Feyereisen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Woodchip ashing protocol</div></div> [STEPS] ?. Place up to 5 crucibles into the muffle furnace ?. Move the muffle furnace near the hood in 197 Borlaug and turn on the hood ?. Turn on the muffle furnace, set ...
["Place up to 5 crucibles into the muffle furnace", "Move the muffle furnace near the hood in 197 Borlaug and turn on the hood", "Turn on the muffle furnace, set temperature to 300o C, leave for 1 hour", "Turn the muffle furnace temperature to 550o C and leave for 4 hours", "After 4 hours, turn off the muffle furnace a...
90,592
Environmental DNA (eDNA) extraction using Qiagen DNeasy 96 Blood and Tissue Kit
1
null
https://www.protocols.io/view/environmental-dna-edna-extraction-using-qiagen-dne-c4p8yvrw
Kathleen Pitz, jbaker, truelove
TITLE: Environmental DNA (eDNA) extraction using Qiagen DNeasy 96 Blood and Tissue Kit AUTHORS: Kathleen Pitz, jbaker, truelove [DESCRIPTION] This protocol is a modified version of the Qiagen DNeasy 96-sample protocol: Purification of Total DNA from Animal Tissues. [STEPS] SECTION: MIOP: Minimum Information about an ...
["[MIOP: Minimum Information about an Omics Protocol] MIOP Term\nValue\n \nmethodology category\n\n \nproject\n\n \npurpose\n\n \nanalyses\n\n \ngeographic location\n\n \nbroad-scale environmental context\n\n \nlocal environmental context\n\n \nenvironmental medium\n\n \ntarget\n\n \ncreator\n\n \nmaterials required\...
null
null
null
dx.doi.org/10.17504/protocols.io.u3peymn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Purpose: Dry Eye Disease (DED) is part of several conditions, including Sjögren’s syndrome (SS) and no single test to diagnosis DED. The present study intends to evaluate whether a set of signs and symptoms of DED can distinguish: a) SS from other non-overlapping systemic diseas...
[]
39,038
Using sequins with RNA sequencing.
1
null
https://www.protocols.io/view/using-sequins-with-rna-sequencing-bic6kaze
Tim Mercer
TITLE: Using sequins with RNA sequencing. AUTHORS: Tim Mercer [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">Sequins are synthetic RNA controls that that are ‘spiked-in’ to your RNA sample, and undergo concurrent library preparation, sequencing and ...
["[Laboratory Steps]\nReceiving sequins.Upon receipt of RNA sequins, first check to ensure they have not thawed during shipment. Please contact us if you have any concerns. Immediately transfer the RNA sequins to frozen storage at -80°C (sequins should not be stored in a -20°C frost-free freezer). Each tube contains RN...
106,655
Purification of GST-TEX264
0
dx.doi.org/10.17504/protocols.io.q26g7178qgwz/v1
https://www.protocols.io/view/purification-of-gst-tex264-dkd74s9n
Elias Adriaenssens
TITLE: Purification of GST-TEX264 AUTHORS: Elias Adriaenssens [DESCRIPTION] This protocol details the purification and analysis of GST-TEX264. [STEPS] SECTION: Purification procedure 1. To purify GST-TEX264, fuse the cytosol-exposed domain of TEX264 (28-313aa) to a N-terminal synthesize gene GST-tag by Genscript and...
["[Purification procedure] To purify GST-TEX264, fuse the cytosol-exposed domain of TEX264 (28-313aa) to a N-terminal synthesize gene GST-tag by Genscript and clone into a pGEX-4T1 vector. Plasmid also available from Addgene.", "[Purification procedure] Collect the supernatant and incubate with pre-equilibrated Glutath...
31,274
Tissue collection and extractions for RNA-seq
null
dx.doi.org/10.17504/protocols.io.basiiece
null
David Lowry
TITLE: Tissue collection and extractions for RNA-seq AUTHORS: David Lowry [STEPS] ?. Pour liquid nitrogen into a polystyrene cooler or another container that can safely hold liquid nitrogen. ?. Remove tissue from plant and place in a tube. Make sure to clean scissors or foreceps with ethanol prior to collections. ?. I...
["Pour liquid nitrogen into a polystyrene cooler or another container that can safely hold liquid nitrogen.", "Remove tissue from plant and place in a tube. Make sure to clean scissors or foreceps with ethanol prior to collections.", "Immediately plunge tube into liquid nitrogen (transcription profiles change rapidly, ...
101,754
BAF_S04_LABCONCO freeze dryer 
0
dx.doi.org/10.17504/protocols.io.261ge5r87g47/v1
https://www.protocols.io/view/baf-s04-labconco-freeze-dryer-dfk23kye
Nicholas Sherman
TITLE: BAF_S04_LABCONCO freeze dryer  AUTHORS: Nicholas Sherman [DESCRIPTION] General operation of the Lyophilizer unit. [BEFORE_START] THIS CHECKLIST SHOULD BE FOLLOWED PRIOR TO EACH USE: 1. remove the accessory drying chamber or manifold from the collector chamber lid and using a soft, lint free paper towel, wipe t...
["[Add samples] MAKE SURE ALL SAMPLES ARE PRE-FROZEN", "[Add samples] Connect a pre-frozen sample to a sample valve on the drying chamber or manifold using an adapter. Turn the plastic valve knob to the “VACUUM” position to open the valve, which connects the attached sample to system vacuum. The bevel on the knob shoul...
58,091
Catalase test for bacterial identification
4
dx.doi.org/10.17504/protocols.io.b4yjqxun
https://www.protocols.io/view/catalase-test-for-bacterial-identification-b4yjqxun
lydiariver
TITLE: Catalase test for bacterial identification AUTHORS: lydiariver [DESCRIPTION] The catalase enzyme is found in most aerobic and facultative anaerobic bacteria that contain cytochrome, being the main exception Streptococcus. Organisms that do not have the cytochrome system also lack the enzyme catalase and there...
["Transfer 2-4 colonies of the bacterial growth in blood agar plates to a glass\nslide using a sterile plastic loop, make circles, and let it dry.", "Place a drop of hydrogen peroxide (H 2 0 2 ) on the glass slide with an\neyedropper.", "Observe immediate results.\nPositive: The oxygen released will be observed as a fo...
99,200
Protocol (C): Automated segmentation of the otic vesicle and image analysis
0
dx.doi.org/10.17504/protocols.io.bp2l6219dgqe/v1
https://www.protocols.io/view/protocol-c-automated-segmentation-of-the-otic-vesi-dc482yzw
Désirée A. Schmitz, Tobias Wechsler, Hongwei Bran Li, Bjoern Menze, Rolf Kümmerli
TITLE: Protocol (C): Automated segmentation of the otic vesicle and image analysis AUTHORS: Désirée A. Schmitz, Tobias Wechsler, Hongwei Bran Li, Bjoern Menze, Rolf Kümmerli [DESCRIPTION] This protocol details the automated segmentation of the otic vesicle and image analysis. [STEPS] SECTION: Automated segmentation ...
["[Automated segmentation of the otic vesicle and image analysis] Organize your images in a folder with two subfolders: \n\none for the brightfield images named ‘Images’, \none for all fluorescence images named ‘Fluor’. \n\nThe image file names have to contain the slice number with a Z as a prefix and the channel numbe...
84,112
Labeling of Zebrafish Organs and Substructures in Light Data in Amira 3D Classic Segmentation Workroom
5
dx.doi.org/10.17504/protocols.io.yxmvm3xobl3p/v1
https://www.protocols.io/view/labeling-of-zebrafish-organs-and-substructures-in-cwdqxa5w
Alyson Petruncio, Aparna Dev, Virginia Ruetten, Gudrun Ihrke, Aubrey Weigel, Project Technical Resources, CellMap Project Team
TITLE: Labeling of Zebrafish Organs and Substructures in Light Data in Amira 3D Classic Segmentation Workroom AUTHORS: Alyson Petruncio, Aparna Dev, Virginia Ruetten, Gudrun Ihrke, Aubrey Weigel, Project Technical Resources, CellMap Project Team [DESCRIPTION] Intended for use on light microscopy datasets of larval zeb...
["[Amira Project and Segmentation Setup] After opening a new Amira project, select File > Open Data, or click+drag the raw data tiff to the empty space under the Project tab. Confirm coordinates and voxel size in the Image Read Parameters window. \n\nFor higher label resolution: Select the arrow of the raw data object ...
null
null
null
dx.doi.org/10.17504/protocols.io.fkvbkw6
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>9 Hourly Timepoints: 0, 1, 2, 3, 4...
[]
23,757
Creatinine Companion Protocol Assay (Exocell)
null
dx.doi.org/10.17504/protocols.io.3fmgjk6
null
Kathi Burke (Frank Brosius lab), Peter Reifsnyder (Ed Leiter Lab)
TITLE: Creatinine Companion Protocol Assay (Exocell) AUTHORS: Kathi Burke (Frank Brosius lab), Peter Reifsnyder (Ed Leiter Lab) [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This assay is designed for companion use w...
["[Urine Collection Protocols]\nSpot UrineBetween 8:00 and 10:00 a.m., the mouse is picked up and induced to urinate into a 1.5 ml microfuge tube. If this is not successful, or not enough is collected, attempts on successive days can be combined (urine should be stored at 4ºC between attempts). Another way to collect s...
49,720
Sample preparation protocol for targeted mass spectrometry analysis of pRabs, total Rabs, LRRK2-pS910, pS935 and total LRRK2
4
dx.doi.org/10.17504/protocols.io.busynwfw
https://www.protocols.io/view/sample-preparation-protocol-for-targeted-mass-spec-busynwfw
Raja S. Nirujogi, Dario Alessi
TITLE: Sample preparation protocol for targeted mass spectrometry analysis of pRabs, total Rabs, LRRK2-pS910, pS935 and total LRRK2 AUTHORS: Raja S. Nirujogi, Dario Alessi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Leucine rich repeat kinase2 (LRRK2) is one of the most commonly implicated kinas...
["[Lysate preparation from mice tissues]\nSnap freeze mouse tissue immediately after isolation in liquid nitrogen and store at .\n-80 °C", "[Lysate preparation from mice tissues]\nPulverize frozen tissue in liquid nitrogen to a fine powder and snap freeze immediately in 2ml tubes. This is done using cell crusher kit.\n...
26,999
PCR-based assay for genotyping of the slick mutation in cattle
null
dx.doi.org/10.17504/protocols.io.6kxhcxn
null
Elizabeth Jannaman, M. Sofia Ortega, Tad S. Sonstegard, Peter J. Hansen
TITLE: PCR-based assay for genotyping of the slick mutation in cattle AUTHORS: Elizabeth Jannaman, M. Sofia Ortega, Tad S. Sonstegard, Peter J. Hansen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Several mutations exist in the prolactin receptor gene (PRLR) of cattle that cause a short, sleek hai...
[]
35,371
Artificial Cerebrospinal Fluid V (ACSF.V)
null
dx.doi.org/10.17504/protocols.io.besjjecn
null
Allen Institute for Brain Science
TITLE: Artificial Cerebrospinal Fluid V (ACSF.V) AUTHORS: Allen Institute for Brain Science [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes how to prepare Artificial Cerebrospinal Fluid (V). Artificial Cerebrospinal Fluid (V) is used during surgical procedures to maintain tis...
[]
42,050
Nuclei prep for single cell RNA/ATAC seq from intestinal surgical samples
4
dx.doi.org/10.17504/protocols.io.bmbak2ie
https://www.protocols.io/view/nuclei-prep-for-single-cell-rna-atac-seq-from-inte-bmbak2ie
Ran Zhou, Oni Basu
TITLE: Nuclei prep for single cell RNA/ATAC seq from intestinal surgical samples AUTHORS: Ran Zhou, Oni Basu [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Isolating high-quality nuclei from intestinal surgical tissues is critical for single cell RNA/ATAC-seq. This protocol provides details on nucl...
["[Pre-processing of surgical samples]\nRinse surgical samples in ice-cold PBS for three times.", "[Pre-processing of surgical samples]\nPlace the surgical samples in a 100 mm petri dish with cold HBSS, and isolate mucosa layer by Iris scissors.", "[Pre-processing of surgical samples]\nWeigh mucosa layer and cut the ti...
null
null
null
dx.doi.org/10.17504/protocols.io.cdhs35
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Protocol for Dephosphorylation of 5&acute;-ends of DNA using CIP in Restriction Enzyme Reaction. Uses the Calf Intestinal&nbsp;Alkaline Phosphatase &nbsp;(CIP - M0290) [STEPS] ?. ?. ?. ?. ?. ?.
[]
36,430
Bulk RNA - Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760, E7765)
1
dx.doi.org/10.17504/protocols.io.bftnjnme
https://www.protocols.io/view/bulk-rna-protocol-for-use-with-nebnext-poly-a-mrna-bftnjnme
Aaron Horning, New England Biolabs
TITLE: Bulk RNA - Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760, E7765) AUTHORS: Aaron Horning, New England Biolabs [DESCRIPTION] The NEBNext Ultra II Directional RNA Library Prep Kit for Illumina contains t...
["[Probe Hybridization to RNA] Prepare the First Strand Synthesis Reaction Buffer and Random Primer Mix (2X) in a nuclease-free microcentrifuge tube as follows: \n\n \nYou can prepare the first strand synthesis reaction buffer later in the protocol, but it is important that it is ready before the elution in step 38. Th...
74,383
Collections Standard Operating Protocol, Plant group: Bryophytes
1
null
https://www.protocols.io/view/collections-standard-operating-protocol-plant-grou-ckvpuw5n
Laura L Forrest, Michelle Hart, David Bell
TITLE: Collections Standard Operating Protocol, Plant group: Bryophytes AUTHORS: Laura L Forrest, Michelle Hart, David Bell [DESCRIPTION] This is part of the collection "DToL Taxon-specific Standard Operating Procedure (SOP) for the Plant Working Group". The SOP collection contains guidance on how to process the vario...
[]
94,278
SOP for DSS kinetics studies
4
dx.doi.org/10.17504/protocols.io.261gedbkdv47/v1
https://www.protocols.io/view/sop-for-dss-kinetics-studies-c8bezsje
Malu G Tansey
TITLE: SOP for DSS kinetics studies AUTHORS: Malu G Tansey [DESCRIPTION] SOP for DSS kinetics studies [STEPS] SECTION: PROTOCOL: 2. Weigh each mouse from a cage individually in small boxes. Record their weights in grams to the nearest tenth for that day. SECTION: PROTOCOL: 3. Keep each mouse in their own box until st...
["[PROTOCOL:] Weigh each mouse from a cage individually in small boxes. Record their weights in grams to the nearest tenth for that day.", "[PROTOCOL:] Keep each mouse in their own box until stools are passed. For a healthy mouse, this should occur quickly. A mouse can be scruffed to expedite this if needed.", "[PROTOC...
54,557
HCR RNA-FISH protocol for the whole-mount brains of Drosophila and other insects
4
dx.doi.org/10.17504/protocols.io.bzh5p386
https://www.protocols.io/view/hcr-rna-fish-protocol-for-the-whole-mount-brains-o-bzh5p386
Amanda A. G. Ferreira, Bogdan Sieriebriennikov, Hunter Whitbeck
TITLE: HCR RNA-FISH protocol for the whole-mount brains of Drosophila and other insects AUTHORS: Amanda A. G. Ferreira, Bogdan Sieriebriennikov, Hunter Whitbeck [DESCRIPTION] This is a protocol to perform RNA fluorescent in situ hybridization (RNA-FISH) using hybridization chain reaction (HCR) on whole-mount samples ...
["[Day 1] Prepare all solutions", "[Day 1] Pre-heat an aliquot of Probe Hybridization Buffer if proceeding with hybridization on the same day (see step case below)\n37 °C", "[Day 1] Dissect brains in cold Schneider's Medium", "[Day 1] Fix brains in 800 μL of Fixation Buffer\n20 min \nRoom temperature\n or", "[Day 1] R...
26,326
Creating a more isogenic strain of worms starting from parental strain
null
dx.doi.org/10.17504/protocols.io.5xwg7pe
null
Cristian Riccio
TITLE: Creating a more isogenic strain of worms starting from parental strain AUTHORS: Cristian Riccio [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes how to create a more isogenic strain starting from a parental strain.</div></div> [STEPS] ?. Pick a single worm into a new f...
["Pick a single worm into a new fresh agar NGM plate. Let it move away from the point of deposition. Pick it again into a new fresh plate. This procedure of first picking into a plate and then into another makes sure that no other worms or eggs are co-picked with the single worm of interest.", "Let the single worm po...
null
null
null
dx.doi.org/10.17504/protocols.io.fcsbiwe
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
20,468
Thawing iPSC Plate
null
dx.doi.org/10.17504/protocols.io.x8ufrww
null
Celeste Karch, Rita Martinez, Jacob Marsh
TITLE: Thawing iPSC Plate AUTHORS: Celeste Karch, Rita Martinez, Jacob Marsh [STEPS] ?. Determine which clones to expand based on screening. ?. Coat 24 well plate with 250-300 ul Matrigel per well (1 well per well, from 96 well). ?. Incubate at for . ?. Remove Styrofoam box from and remove plate. Check on plate afte...
["Determine which clones to expand based on screening.", "Coat 24 well plate with 250-300 ul Matrigel per well (1 well per well, from 96 well).", "Incubate at for .", "Remove Styrofoam box from and remove plate.\nCheck on plate after to avoid over-thawing.", "To each well add mTesR1 supplemented with 10 uM Rock inh...
25,586
06 Gel purification
null
dx.doi.org/10.17504/protocols.io.48sgzwe
null
TJUSLS China
TITLE: 06 Gel purification AUTHORS: TJUSLS China [STEPS] ?. Column balancing step: add 500ul buffer BL to the adsorption column CA2 (the adsorption column is put into the collection tube), centrifuge at 12,000 RPM (~13,400×g) for 1 min, dump the waste liquid in the collection tube, and put the adsorption column back i...
["Column balancing step: add 500ul buffer BL to the adsorption column CA2 (the adsorption column is put into the collection tube), centrifuge at 12,000 RPM (~13,400×g) for 1 min, dump the waste liquid in the collection tube, and put the adsorption column back into the collection tube.\n500 µl", "Remove the single targe...
98,428
Parse Evercode WT v2 -- University of Minnesota TMCs
0
dx.doi.org/10.17504/protocols.io.dm6gpz8rplzp/v2
https://www.protocols.io/view/parse-evercode-wt-v2-university-of-minnesota-tmcs-dcc42syw
Laura J Niedernhofer, David A Bernlohr
TITLE: Parse Evercode WT v2 -- University of Minnesota TMCs AUTHORS: Laura J Niedernhofer, David A Bernlohr [DESCRIPTION] Combinatorial barcoding is an instrument-free method used to generate single-cell and single-nucleus RNA-sequencing libraries. Outlined here are the methods used to isolate single-nuclei from froze...
["[Single Nuclei Dissociation]", "[Nuclei Fixation Protocol]", "[Evercode WT Protocol]", "[FASTQ Generation] BCL data from Illumina sequencer is demultiplexed and converted into FASTQ format using bcl2fastq version 2.20.0. \n\nMultiplexed FASTQ files were split by sample using a script provided by Parse Biosciences (pa...
44,555
Quality of Care Index (QCI)
5
dx.doi.org/10.17504/protocols.io.bprjmm4n
https://www.protocols.io/view/quality-of-care-index-qci-bprjmm4n
Esmaeil Mohammadi, Erfan Ghasemi, Sahar Saeedi Moghaddam, Moein Yoosefi, Shahin Roshani, Naser Ahmadi, Masoud Masinaei, Maryam Nasserinejad, Narges Ebrahimi, Sina Azadnajafabad, Mahtab Rouhifard Khalilabad, Negar Rezaei, Ali H Mokdad, Bagher Larijani, Farshad Farzadfar
TITLE: Quality of Care Index (QCI) AUTHORS: Esmaeil Mohammadi, Erfan Ghasemi, Sahar Saeedi Moghaddam, Moein Yoosefi, Shahin Roshani, Naser Ahmadi, Masoud Masinaei, Maryam Nasserinejad, Narges Ebrahimi, Sina Azadnajafabad, Mahtab Rouhifard Khalilabad, Negar Rezaei, Ali H Mokdad, Bagher Larijani, Farshad Farzadfar [DESC...
["[Data acquisition]\nA. Consider retrieving data from the GBD data repository [https://vizhub.healthdata.org/gbd-compare/].B. Use the left sidebars and menus to navigate for causes.C. Primary parameters are required 1. age-standardized mortality rate 2. age-standardized prevalence rate 3. age-standardized incidence ra...
44,724
Cell DIVE™ Platform | Slide Clearing and Antigen Retrieval
4
dx.doi.org/10.17504/protocols.io.bpwumpew
https://www.protocols.io/view/cell-dive-platform-slide-clearing-and-antigen-retr-bpwumpew
Liz McDonough, Chrystal Chadwick, Fiona Ginty, Christine Surrette, Anup Sood
TITLE: Cell DIVE™ Platform | Slide Clearing and Antigen Retrieval AUTHORS: Liz McDonough, Chrystal Chadwick, Fiona Ginty, Christine Surrette, Anup Sood [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The purpose of this protocol is to manually deparaffinize and rehydrate slides for the Cell DIVE™ Pl...
["[Slide Baking]\nThis is the initial step of the deparaffinization process. This step serves two purposes: (1) melting of the paraffin wax and (2) ensuring tissue adherence to the slides.", "[Slide Baking]\nSet the incubator to .\n60 °C", "[Slide Baking]\nLoad tissue slides into a slide rack. All tissue slides should ...
41,852
Freeze-drying (Lyophilization) of CoronaDetective tubes
4
dx.doi.org/10.17504/protocols.io.bk44kyyw
https://www.protocols.io/view/freeze-drying-lyophilization-of-coronadetective-tu-bk44kyyw
Guy Aidelberg, Rachel Aronoff
TITLE: Freeze-drying (Lyophilization) of CoronaDetective tubes AUTHORS: Guy Aidelberg, Rachel Aronoff [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A protocol for freeze-drying (Lyophilization) of CoronaDetective tests, </div><div class = "text-block">and more generally any QUASR RT-LAMP reaction....
["Thaw components (dNTPs, 10X Primer mixes (need to be made), MgSO4, Isothermal amplification buffer, and Enzymes)Vortex and quickly spin tubes down before opening for dispensing.This protocol is for one standard 96 well PCR plate and can be scaled as needed.", "10X Primer mix: assuming your primer stocks are at for...
29,193
WTc11 iPSC Culture and Maintenance
null
dx.doi.org/10.17504/protocols.io.8rhhv36
null
Connor Ludwig, Ruilin Tian, Martin Kampmann
TITLE: WTc11 iPSC Culture and Maintenance AUTHORS: Connor Ludwig, Ruilin Tian, Martin Kampmann [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol explains general culture and maintenance of the WTc11 iPSC cell line. </div></div> [STEPS] ?. [Prepare complete StemFlex Medium] Thaw the froz...
["[Prepare complete StemFlex Medium]\nThaw the frozen StemFlex Supplement 10X at for ~ or overnight at to .\n0 Room temperature\n2 °C\n8 °C\nIMPORTANT! Do not thaw the frozen supplement at 37°C.", "[Prepare complete StemFlex Medium]\nMix the thawed supplement by gently inverting 3−5 times.", "[Prepare complete StemFl...
87,394
Staining of CD31 and CD13 in PDGFR-B/Td-tomato brain sections
1
dx.doi.org/10.17504/protocols.io.x54v9p2b1g3e/v1
https://www.protocols.io/view/staining-of-cd31-and-cd13-in-pdgfr-b-td-tomato-bra-czkax4se
Daniel Manrique-Castano
TITLE: Staining of CD31 and CD13 in PDGFR-B/Td-tomato brain sections AUTHORS: Daniel Manrique-Castano [DESCRIPTION] This protocol is suitable for the staining of CD31 and CD13 in PDGFR-B/Td-Tomato fixed mouse brain sections. [GUIDELINES] Read the entire protocol before starting the procedure. Note that this protocol...
["[Tissue preparation and blocking] Extract the sections from the anti-freeze media and rinse them in PBS using a 24-well plate.", "[Tissue preparation and blocking] Aspirate the PBS and incubate the sections in the Blocking solution for 60 min at Room temperature .", "[Antibody incubation] When blocking is finished, a...
99,877
10% Tween 20
1
dx.doi.org/10.17504/protocols.io.ewov18m7pgr2/v2
https://www.protocols.io/view/10-tween-20-ddsd26a6
Allen Institute for Brain Science
TITLE: 10% Tween 20 AUTHORS: Allen Institute for Brain Science [DESCRIPTION] This protocol is used to prepare 10% Tween 20. Tween 20, when added to aqueous solutions acts as a surfactant, decreasing the surface tension of the solution. It also punches holes in membranes, allowing easier penetration of macromolecules i...
[]
48,181
MojoSort™ Pan DC Isolation Kit Column Protocol
4
null
https://www.protocols.io/view/mojosort-pan-dc-isolation-kit-column-protocol-btavnie6
Ken Lau
TITLE: MojoSort™ Pan DC Isolation Kit Column Protocol AUTHORS: Ken Lau [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">MojoSort™ Pan DC Isolation Kit Column Protocol</span><span></span></div></div> [STEPS] ?. [Protocol] Prepare cells from your tissue of interest or...
["[Protocol]\nPrepare cells from your tissue of interest or blood without lysing erythrocytes.", "[Protocol]\nIn 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."...
63,322
DNA extraction (BOMB)
4
dx.doi.org/10.17504/protocols.io.x54v9ywq1g3e/v1
https://www.protocols.io/view/dna-extraction-bomb-b932r8qe
Tsu-Chun Hung, Yin-Tse Huang
TITLE: DNA extraction (BOMB) AUTHORS: Tsu-Chun Hung, Yin-Tse Huang [DESCRIPTION] DNA extraction (BOMB) [STEPS] SECTION: Sample Collection 1. Add 200 µL of 1mm beads to 1.5ml enppendorf tube SECTION: Sample Collection 2. Add200 µL of 0.5mm beads to 1.5ml enppendorf tube SECTION: Sample Collection 3. Add 225 µL o...
["[Sample Collection] Add 200 µL of 1mm beads to 1.5ml enppendorf tube", "[Sample Collection] Add200 µL of 0.5mm beads to 1.5ml enppendorf tube", "[Sample Collection] Add 225 µL of TE buffer to 1.5ml enppendorf tube", "[Sample Collection] Add 375 µL of lysis buffer to 1.5ml enppendorf tube", "[Sample Collection] Collec...
16,806
Data File
null
dx.doi.org/10.17504/protocols.io.uneevbe
null
Yolonda Freeman-Hildreth
TITLE: Data File AUTHORS: Yolonda Freeman-Hildreth [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Data file; </div><div class = "text-block">Total sample population</div><div class = "text-block">Facebook/Social connection group</div><div class = "text-block">Amazon Mechanical Turk group</div><div...
[]
85,513
GONE in 360 seconds: gentleMACS Octo Dissociator-based Nuclei Extraction
4
dx.doi.org/10.17504/protocols.io.dm6gp3bo8vzp/v1
https://www.protocols.io/view/gone-in-360-seconds-gentlemacs-octo-dissociator-ba-cxrhxm36
Deepak Balamurali, Kinga M. Wrona, Lea Eich, Anika Witten, Alba M. Albert, Marc N. Hirt, Thomas Eschenhagen, Monika Stoll
TITLE: GONE in 360 seconds: gentleMACS Octo Dissociator-based Nuclei Extraction AUTHORS: Deepak Balamurali, Kinga M. Wrona, Lea Eich, Anika Witten, Alba M. Albert, Marc N. Hirt, Thomas Eschenhagen, Monika Stoll [DESCRIPTION] Single-cell and single-nuclei approaches are being extensively used to decipher the cellular h...
["[1. Pre-processing] Pre-cool the buffers, and consumables with sample contact (e.g. gentleMACS C Tube and SmartStrainers) at 4 °C 15 min. Similarly, place the gentleMACS Octo Coolers in -20 °C .", "[2. Nuclei Extraction] Add 2 mL ice-cold lysis buffer to each pre-cooled gentleMACS C Tube.", "[4.\tQuality Check/Cont...
109,383
STREAM Benthic Metabarcoding Lab Protocol
1
dx.doi.org/10.17504/protocols.io.5jyl8ppb6g2w/v2
https://www.protocols.io/view/stream-benthic-metabarcoding-lab-protocol-dn3f5gjn
Carley Maitland, Michael Wright, Mehrdad Hajibabaei
TITLE: STREAM Benthic Metabarcoding Lab Protocol AUTHORS: Carley Maitland, Michael Wright, Mehrdad Hajibabaei [DESCRIPTION] STREAM (Sequencing the Rivers for Environmental Assessment and Monitoring; www.stream-dna.org) is a Canada-wide community-based science program established in 2018 which is led by the research la...
["[Homogenizing] Blenders should be decontaminated ahead of homogenization. Clean by fully disassembling blender components and rinsing with water. Visually inspect to ensure all debris has been removed before scrubbing all surfaces with ELIMINase ™ and rinsing with deionized (DI) water. Treat with UV light for 30 min ...