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68,512
Diagnostic Restriction Digest
4
null
https://www.protocols.io/view/diagnostic-restriction-digest-ce58tg9w
Brian Teague
TITLE: Diagnostic Restriction Digest AUTHORS: Brian Teague [DESCRIPTION] In this restriction digest, you'll use an enzyme that cuts DNA to cut your miniprepped plasmid. This can give you some evidence as to whether your plasmid is what you expected or not. You'll also use Benchling to predict the result of you...
["[Perform the diagnostic digest] For each miniprep, compute the volume that contains 1 µg of DNA.", "[Perform the diagnostic digest] In the PCR tube, mix:\nThe volume of DNA you computed in step 1, up to a maximum of 5 µL\n2 µL of CutSmart enzyme buffer\n2 µL of PvuII enzyme\nEnough nuclease-free water for a total vol...
null
null
null
dx.doi.org/10.17504/protocols.io.rswd6fe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div> <div> <div> <div> <div> <div> <div> <div> <div> <div> <div> <div> <div> <p>The protocol aims explicitly to amplify BFV viruses and not other viruses.</p> <p>The assay targets the E2 gene region and is designed as a qualitative test for investigating BFV infection of humans...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.pifdkbn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Here is a short intro on short read assembly using the SPAdes assembler. </p> <p> </p> <p>Some notes on tool installation:</p> <p>To install SPAdes or other tools into your PATH you may wish to use a package manager called Miniconda. For this go to the website here and downlo...
[]
63,833
Via Keto Gummies Expert Reviews – Official Report Analysis
1
dx.doi.org/10.17504/protocols.io.81wgb69eolpk/v1
https://www.protocols.io/view/via-keto-gummies-expert-reviews-official-report-an-cajzscp6
D D
TITLE: Via Keto Gummies Expert Reviews – Official Report Analysis AUTHORS: D D [DESCRIPTION] This is definitively what the new and progressiveVia Keto Gummiesis about. It's a ketogenic supplement planned explicitly to assist you with arriving at ketosis faster. At the end of the day, the ideal answer for anybody need...
[]
25,247
How to Prepare a Single Cell Suspension from Mouse Spleen
null
dx.doi.org/10.17504/protocols.io.4v7gw9n
null
STEMCELL Technologies
TITLE: How to Prepare a Single Cell Suspension from Mouse Spleen AUTHORS: STEMCELL Technologies [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The spleen is an important organ of the immune system responsible for filtering blood and initiating immune responses to blood-borne antigens. Various blood...
["[Mechanical Digestion of a Spleen Sample]\nTransfer the spleen to be processed into a sterile 35 mm culture dish containing 5 mL of recommended dissociation medium for downstream isolation. If you are not using an isolation kit following dissociation please use phosphate buffered saline (PBS) + 1 mM EDTA.Recommended ...
88,915
Simulating the modal analysis of hyperelastic membranes immersed in fluid using FE software ANSYS
5
dx.doi.org/10.17504/protocols.io.bp2l6x7dklqe/v2
https://www.protocols.io/view/simulating-the-modal-analysis-of-hyperelastic-memb-c23tygnn
samuel.vorlet
TITLE: Simulating the modal analysis of hyperelastic membranes immersed in fluid using FE software ANSYS AUTHORS: samuel.vorlet [DESCRIPTION] This protocol provides step-by-step guidelines to perform the modal analysis of pre-strained hyperelastic rectangular membrane accounting for fluid-structure interactions using ...
["[Hyperelastic material definition using the Mooney-Rivlin formulation from uniaxial test data] Open Engineering Data.", "Ad a new material and define the material name. Verify the units.", "Define the hyperelastic material properties.", "In the Hyperelastic toolbox, choose the Mooney-Rivlin material model with the ap...
104,376
Sinai SCENT TMC - FFPE Blocking, Sectioning, and TMA Construction
0
null
https://www.protocols.io/view/sinai-scent-tmc-ffpe-blocking-sectioning-and-tma-c-dh6y39fw
Sojin Kim
TITLE: Sinai SCENT TMC - FFPE Blocking, Sectioning, and TMA Construction AUTHORS: Sojin Kim [DESCRIPTION] FFPE Blocking and Sectioning protocol [GUIDELINES] Comply with Universal Precautions when handling all specimens. Use personal protective equipment according to the institution’s guidelines. [STEPS] SECTION: FF...
["[FFPE Blocking and Sectioning] Rince the collected tissues in PBS to remove blood.", "[FFPE Blocking and Sectioning] Place tissues in at least 10 volumes of buffered formalin or buffered paraformaldehyde", "[FFPE Blocking and Sectioning] Incubate for the necessary fixation time\n1. 1-2 mm thick: 2-3 hours RT\n2. 5-10...
null
null
null
dx.doi.org/10.17504/protocols.io.ruhd6t6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This assay is used to measure cell viability. Dead cells have damage membranes. The ethidium homodimer-1 enters damaged cells and is fluorescent when bound to nucleic acids, producing a bright red fluorescence in damaged or dead cells.</p> [STEPS]
[]
72,965
Untargeted lipidomics analysis for Golgi immunopurification (Golgi-IP)
4
dx.doi.org/10.17504/protocols.io.3byl4jq6jlo5/v1
https://www.protocols.io/view/untargeted-lipidomics-analysis-for-golgi-immunopur-cjhduj26
Wentao Dong, Eshaan S Rawat, Monther Abu-Remaileh
TITLE: Untargeted lipidomics analysis for Golgi immunopurification (Golgi-IP) AUTHORS: Wentao Dong, Eshaan S Rawat, Monther Abu-Remaileh [DESCRIPTION] The Golgi apparatus functions as a central hub in the cell that processes, packages, and distributes proteins. Despite its critical cellular function, there has been ch...
["[LC/MS lipidomics settings] Set an ID-X tribrid mass spectrometer (Thermo Fisher Scientific) with a heated electrospray ionization (HESI) probe, for initial nonpolar lipid profiling.\n\nPrepare an Ascentis Express C18 150 x 2.1 mm column (Millipore Sigma 53825-U) coupled with a 5 x 2.1 mm guard (Sigma-Aldrich 53500-U...
24,556
Hiseq 2000 Library Construction and Sequencing for RNA Seq
null
dx.doi.org/10.17504/protocols.io.38kgruw
null
Eric J. Carpenter, Naim Matasci, Shuangxiu Wu, Jing Sun, Jun Yu, Fabio Rocha Jimenez Vieira, Chris Bowler, Richard G. Dorrell, Matt Gitzendanner, Ling Li, Wensi Du, Kristian Ullrich, Michael S. Barker, James H. Leebens-Mack, Gane Ka-Shu Wong
TITLE: Hiseq 2000 Library Construction and Sequencing for RNA Seq AUTHORS: Eric J. Carpenter, Naim Matasci, Shuangxiu Wu, Jing Sun, Jun Yu, Fabio Rocha Jimenez Vieira, Chris Bowler, Richard G. Dorrell, Matt Gitzendanner, Ling Li, Wensi Du, Kristian Ullrich, Michael S. Barker, James H. Leebens-Mack, Gane Ka-Shu Wong [D...
["Isolate polyA RNA from of total RNA treated by using .It is best to use up to 50 µg as the use of a lower mass (typically 20 µg) has been insufficient for successful library construction. This can be assessed by running final PCR products on an agarose gel; the library construction is considered to have failed when...
71,727
Rapid Sequencing gDNA
1
dx.doi.org/10.17504/protocols.io.14egn27eyg5d/v1
https://www.protocols.io/view/rapid-sequencing-gdna-ciapuadn
Carlos Goller
TITLE: Rapid Sequencing gDNA AUTHORS: Carlos Goller [DESCRIPTION] ONT Rapid sequencing kit use in a classroom setting. [STEPS] SECTION: Library Preparation 1. DNA tagmentation Thaw kit components at Room temperature , spin down briefly using a microfuge and mix by pipetting as indicated below: Lambda DNA (50 μg/ml...
["[Library Preparation] DNA tagmentation\n\nThaw kit components at Room temperature , spin down briefly using a microfuge and mix by pipetting as indicated below:\n\nLambda DNA (50 μg/ml): thaw at RT, briefly spin down, mix well by pipetting\nFragmentation Mix (FRA): not frozen, briefly spin down, mix well by pipetting...
44,042
Vivarium Population Spenser: Internal migration module
5
dx.doi.org/10.17504/protocols.io.bn9imh4e
https://www.protocols.io/view/vivarium-population-spenser-internal-migration-mod-bn9imh4e
Camila Rangel Smith, Kasra Hosseini
TITLE: Vivarium Population Spenser: Internal migration module AUTHORS: Camila Rangel Smith, Kasra Hosseini [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Description of the steps followed by Vivarium Population Spenser library when running the Internal migration module. </div></div> [STEPS] ?. Thi...
["This module models the [internal_migration](src/vivarium_population_spenser/internal_migration.py) between MSOAs (and theirrespective LADs) of individuals based on their gender, age, initial location (local authority level) and ethnicity.The input table to establish the pool of migrants that internally migrate based...
40,798
ELISA for quantification of IL-34 in human serum.
6
dx.doi.org/10.17504/protocols.io.bj36kqre
https://www.protocols.io/view/elisa-for-quantification-of-il-34-in-human-serum-bj36kqre
Angel Justiz-Vaillant
TITLE: ELISA for quantification of IL-34 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-34 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-34 present in the serum sample binds to antibodies adsorbed into the microwells.", "The microplate is blocked with 3% non-fat milk-PBS buffer and...
107,221
Multicellular Circulating Co-Culture
0
dx.doi.org/10.17504/protocols.io.ewov19b47lr2/v1
https://www.protocols.io/view/multicellular-circulating-co-culture-dkxv4xn6
Bianca Cruz Pachane, Pedro Henrique Teixeira Bottaro, Wanessa Fernanda Altei, Heloisa Sobreiro Selistre de Araujo
TITLE: Multicellular Circulating Co-Culture AUTHORS: Bianca Cruz Pachane, Pedro Henrique Teixeira Bottaro, Wanessa Fernanda Altei, Heloisa Sobreiro Selistre de Araujo [DESCRIPTION] A novel method to study the tumor microenvironment (TME) in vitro, using the quasi-vivo technology from Kirstall to survey the individual ...
["[Preparation of matrix-coated coverslips] Clean round glass coverslips (13 mm ø) with 70% ethanol wipes before use. Maintain slips in a clean container.", "[Preparation of matrix-coated coverslips] Prepare a 0.5% solution of glutaraldehyde in H2O and maintain it at 4 °C protected from light.", "[Preparation of matrix...
null
null
null
dx.doi.org/10.17504/protocols.io.mpqc5mw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
80,233
Generation of stable cell lines using retroviral system
4
dx.doi.org/10.17504/protocols.io.81wgbyez1vpk/v1
https://www.protocols.io/view/generation-of-stable-cell-lines-using-retroviral-s-cskhwct6
nguyen.tha
TITLE: Generation of stable cell lines using retroviral system AUTHORS: nguyen.tha [DESCRIPTION] This protocol details generation of stable cell lines using retroviral system. [GUIDELINES] Attention The HEK293T cells detach very easily, be extra gentle when changing the media. [STEPS] SECTION: Day 1 1. Seed NIH HE...
["[Day 1] Seed NIH HEK293T cells into a 6-well plate (900k cells/well if set up in the morning, 950k cells/well if set up in the afternoon).", "[Day 2: The following protocol is designed for one well of the 6-well plate] Transfect cells with viral and helper vectors using lipofectamine LTX. Combine the following in a 1...
88,854
PCR based amplicon sequencing of P. vivax antigens
4
dx.doi.org/10.17504/protocols.io.261gedwddv47/v1
https://www.protocols.io/view/pcr-based-amplicon-sequencing-of-p-vivax-antigens-c2zwyf7e
Paolo Bareng
TITLE: PCR based amplicon sequencing of P. vivax antigens AUTHORS: Paolo Bareng [STEPS] SECTION: Primer pool(s) preparation 1. Prepare the primer pools 1, 2, and 3 by reconstituting lyophilized primers to a concentration of 100µM using nuclease-free water SECTION: Primer pool(s) preparation 1.1. The tables below show ...
["[Primer pool(s) preparation] Prepare the primer pools 1, 2, and 3 by reconstituting lyophilized primers to a concentration of 100µM using nuclease-free water", "[Primer pool(s) preparation] The tables below show the volume of each 100µM forward and reverse primers stock to be added in the respective pools. The total ...
null
null
null
dx.doi.org/10.17504/protocols.io.hbvb2n6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Current long read sequencing, e. g. PacBio and Nanopore, requires high molecular weight (HMW) and highly pure DNA. Many fungal and plant species have high content of polysaccharides and other contaminants that are co-precipitated with DNA during ethanol precipitation. This pr...
[]
64,751
Standard Operating Procedure for assembly and deployment of ovitraps
1
dx.doi.org/10.17504/protocols.io.14egn7r4pv5d/v1
https://www.protocols.io/view/standard-operating-procedure-for-assembly-and-depl-cbgpsjvn
Tanya L Russell, Kyran Staunton, Thomas R. Burkot
TITLE: Standard Operating Procedure for assembly and deployment of ovitraps AUTHORS: Tanya L Russell, Kyran Staunton, Thomas R. Burkot [DESCRIPTION] The purpose of this Standard Operating Procedure (SOP) is to outline the materials and processes required to assemble, deploy and service an ovitrap. Description: Ovit...
["[Trap assembly] Collect trap components. Ovistrips made from cloth or paper should be cut to roughly 6 cm wide x 12 cm long.", "[Trap assembly] Pour water into the bucket until it is about 2/3 full. For example, fill a 1 L bucket with ~660 ml of water.", "[Trap assembly] Insert organic material (0.5 g/L) of such as a...
null
null
null
dx.doi.org/10.17504/protocols.io.r4xd8xn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The Modified Ziehl-Neelsen stain (mZN stain) is a type of differential bacteriological stain used to identify acid-fast organisms, mainly <em>Mycobacteria</em>. Acid fast organisms are those which are capable of retaining the primary stain when treated with an acid (<em>fast=...
[]
34,482
Extracellular DNA extraction
null
dx.doi.org/10.17504/protocols.io.bdwsi7ee
null
Charline Giguet-Covex, Pierre Taberlet, Francesco Gentile Ficetola
TITLE: Extracellular DNA extraction AUTHORS: Charline Giguet-Covex, Pierre Taberlet, Francesco Gentile Ficetola [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Over the past decade, an increasing number of studies has used environmental DNA from lake sediments to trace past lake ecosystem and landsc...
["[exDNA extraction]\nPhosphate buffer preparation:The phosphate buffer must be prepared the same day, before starting the extraction protocol. Calculate the quantity of phosphate buffer required for the extractions. Prepare a little more especially for the extraction control. You will add the same volume of phosphate ...
35,883
Successful and cost-effective DNA extraction method from insects with small amount of tissue
null
dx.doi.org/10.17504/protocols.io.bfajjicn
https://www.protocols.io/view/successful-and-cost-effective-dna-extraction-metho-bfajjicn
Rahul Jamdade
TITLE: Successful and cost-effective DNA extraction method from insects with small amount of tissue AUTHORS: Rahul Jamdade [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">DNA extraction from insects is an initial and critical step that might affect the molecular taxonomy. There are several protoc...
[]
44,640
Dip-C (Part 2: Whole-genome Amplification with Nextera)
4
dx.doi.org/10.17504/protocols.io.bpt8mnrw
https://www.protocols.io/view/dip-c-part-2-whole-genome-amplification-with-nexte-bpt8mnrw
Longzhi Tan
TITLE: Dip-C (Part 2: Whole-genome Amplification with Nextera) AUTHORS: Longzhi Tan [STEPS] ?. [Oligos] Carrier ssDNA (same as in LIANTI and META):TCAGGTTTTCCTGAAPurification: standard desaltingDissolve in 0.1 X TE (made from ) to a final concentration of .Store at . -20 °C ?. [Oligos] Nextera i7 Index Primers:701: C...
["[Oligos]\nCarrier ssDNA (same as in LIANTI and META):TCAGGTTTTCCTGAAPurification: standard desaltingDissolve in 0.1 X TE (made from ) to a final concentration of .Store at .\n-20 °C", "[Oligos]\nNextera i7 Index Primers:701: CAAGCAGAAGACGGCATACGAGATTCGCCTTAGTCTCGTGGGCTCGG702: CAAGCAGAAGACGGCATACGAGATCTAGTACGGTCTCGTG...
50,414
Protocol for Image processing and analysis of VPS13D recruitment to mitochondria
1
dx.doi.org/10.17504/protocols.io.bvgnn3ve
https://www.protocols.io/view/protocol-for-image-processing-and-analysis-of-vps1-bvgnn3ve
Marianna Leonzino, Andrés Guillén-Samander, Ni Tang, Pietro De Camilli
TITLE: Protocol for Image processing and analysis of VPS13D recruitment to mitochondria AUTHORS: Marianna Leonzino, Andrés Guillén-Samander, Ni Tang, Pietro De Camilli [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol details the image processing and analysis of VPS13D recruitment ...
["[For the analysis of optogenetic experiments:]\nBuild kymographs tracing a line ROI across the mitochondria that was illuminated with Blue Light.", "[For the analysis of optogenetic experiments:]\nMeasure an intensity profile by tracing a line ROI across the center of the mitochondrial signal on the kymograph. Take a...
94,189
Sucrose preference test 
4
dx.doi.org/10.17504/protocols.io.3byl4q4xrvo5/v1
https://www.protocols.io/view/sucrose-preference-test-c78mzru6
mariangela.massarocenere
TITLE: Sucrose preference test  AUTHORS: mariangela.massarocenere [DESCRIPTION] The sucrose preference test assesses the animal’s interest in a sweet-tasting sucrose solution relative to unsweetened water [STEPS] 1. Place one animalper cage during the test duration and habituate them to drinking from two equally acce...
["Place one animalper cage during the test duration and habituate them to drinking from two equally accessible bottles", "Refill each bottle with fresh water or 1% sucrose solution and inverte to its original position every day for two consecutive days", "To avoid side preference, place the sucrose bottle on the left f...
null
null
null
dx.doi.org/10.17504/protocols.io.d9m945
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol has been modified from the original method developed and reported by Steward and Culley (2010) and Mueller et al. (2014). As a starting point we assume that viruses have been collected from the environment on 0.02 µm syringe filters (Anotop, Whatman).</p> [STEP...
[]
51,102
Flow CyTOF Using Single Cells From Human Islets
4
dx.doi.org/10.17504/protocols.io.bv56n89e
https://www.protocols.io/view/flow-cytof-using-single-cells-from-human-islets-bv56n89e
Klaus H. Kaestner Lab, Suzanne Shapira
TITLE: Flow CyTOF Using Single Cells From Human Islets AUTHORS: Klaus H. Kaestner Lab, Suzanne Shapira [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Cytometry by time-of-flight (CyTOF) is an application of mass cytometry using antibodies conjugated to rare heavy m...
["[II. CyTOF barcoding and labelling]\nThaw the cells in quickly, and immediately add the Perm bufferRemove the barcodes (Fluidigm 201060) from freezer and allow them to warm up to RT for at least 10 min.\n37 °C\n-20 °C", "[II. CyTOF barcoding and labelling]\nWash cells 2x with Foxp3 perm buffer (eBioscience, 00-5523...
null
null
null
dx.doi.org/10.17504/protocols.io.etxbepn
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <strong>Materials:</strong><br /><br /> <ol> <li>Reagent “A” - 2.0% Na<sub>2</sub>CO<sub>3</sub> in 0.1 N NaOH</li> <li>Reagent “B” - 0.5% CuSO<sub>4</sub><sup>.</sup>5H<sub>2</sub>O in 1.0% Na citrate</li> <li>Reagent “C” - 50.0 mL reagent “A” + 1.0 mL reagent “B”</li> <li>Reage...
[]
59,738
Total RNA and DNA from Microalgae (24 samples per day)
1
null
https://www.protocols.io/view/total-rna-and-dna-from-microalgae-24-samples-per-d-b6j2rcqe
Yingyu Hu, Zoe V Finkel
TITLE: Total RNA and DNA from Microalgae (24 samples per day) AUTHORS: Yingyu Hu, Zoe V Finkel [DESCRIPTION] Here we describe a protocol for extracting and quantifying bulk RNA and DNA from microalgae, which is adapted from Berdalet E. et al. (2005). RNA and DNA are extracted from microalgae samples and then quant...
["[Day 1: Freeze-dry samples] Freeze dry samples and blank filters. Freeze at -80 °C until processed.", "[Day 1: Prepare primary solutions] Turn on UV light in biosafety cabinet for 15 min and clean working surface with decontamination solution.", "[Day 1: Prepare primary solutions] Prepare Tris buffer 5 mM pH 8.0", ...
23,764
CODEX Oligo-labeled Antibody Conjugation
null
dx.doi.org/10.17504/protocols.io.3fugjnw
null
Yury Goltsev, Nikolay Samusik, Julia Kennedy-Darling, Salil Bhate, Matthew Hale, Gustavo Vazquez, Sarah Black, Garry Nolan
TITLE: CODEX Oligo-labeled Antibody Conjugation AUTHORS: Yury Goltsev, Nikolay Samusik, Julia Kennedy-Darling, Salil Bhate, Matthew Hale, Gustavo Vazquez, Sarah Black, Garry Nolan [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>CODEX is a technology that uses oligo labeled antibodies, speciali...
["[Antibody Disulfide Reduction Reaction]\nRetrieve one MWCO filter column for each antibody to be conjugated. Block nonspecific antibody binding to the MWCO filter columns by adding 500ul Filter Blocking Solution to the top of each column and spinning down at 12,000g for 2 minutes.\n0 Room temperature", "[Antibody Di...
40,889
Removal of Melanin
1
dx.doi.org/10.17504/protocols.io.bj6zkrf6
https://www.protocols.io/view/removal-of-melanin-bj6zkrf6
Jason Stajich
TITLE: Removal of Melanin AUTHORS: Jason Stajich [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Principle is that CTAB is charging the anionic nucleotides whereby neutral polysaccharides/ melanins are remaining in supernatant. This method also uses urea with the idea that the presence of urea helps...
["Add to ~- DNA/RNA solution until a volume of is reached.\n100 µl\n200 µl\n400 µl", "Add\n130 µl", "Add mL of CTAB-Urea solution\n1.6 mL\n{\"blocks\":[{\"key\":\"7ul3n\",\"text\":\"\",\"type\":\"unstyled\",\"depth\":0,\"inlineStyleRanges\":[],\"entityRanges\":[],\"data\":[]}],\"entityMap\":[]}", "Mix samples (by han...
61,598
Human fibroblast culturing
4
dx.doi.org/10.17504/protocols.io.rm7vzy542lx1/v1
https://www.protocols.io/view/human-fibroblast-culturing-b8d6rs9e
Laura Smith, David C
TITLE: Human fibroblast culturing AUTHORS: Laura Smith, David C [DESCRIPTION] Fibroblasts are cultured in Dulbecco’s modified eagle media (DMEM) 4500 (mg/L) growth medium supplemented with Glutamax (Gibco), 10% foetal bovine serum (FBS), non-essential amino acids (NEAA: 0.1 mM of: glycine, L-alanine, L-asparagine, L...
["[Characteristics] Characteristics\nObtained /developed from human adult dermal biopsies\nProliferative until p20\nDoubling time ~ 24h\nMaintain between 60 – 90% confluent", "[Complete growth medium] DMEM (LT # 61965-059; 4500 mg/L and no pyruvate) Glutamax medium\nFetal bovine serum (FBS, 10% final) – 1 / 1...
null
null
null
dx.doi.org/10.17504/protocols.io.c3iykd
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Last optimized January 2006, updated by Matt on March 16 2009 [GUIDELINES] This protocol comes from a group of other protocols.This protocol is (1) of (4): <br />1. <a href="https://www.protocols.io/view/Large-Volume-Marine-Cyanophage-Phage-Protocols-c3iykd" target="_blank">'La...
[]
47,834
Modified EMP ITS Illumina Amplicon Protocol 
1
dx.doi.org/10.17504/protocols.io.6qpvrdmeogmk/v1
https://www.protocols.io/view/modified-emp-its-illumina-amplicon-protocol-bsx2nfqe
Dylan P. Smith, Kabir G. Peay, Gail Ackermann, Amy Apprill, Markus Bauer, Donna Berg-Lyons, Jason Betley, T. D. Bruns, J. Greg Caporaso, Noah Fierer, Louise Fraser, Jed A. Fuhrman, M. Gardes, Jack A. Gilbert, Niall Gormley, Greg Humphrey, James Huntley, Janet K. Jansson, Rob Knight, Chris L. Lauber, S. Lee, Sarah M. Ow...
TITLE: Modified EMP ITS Illumina Amplicon Protocol  AUTHORS: Dylan P. Smith, Kabir G. Peay, Gail Ackermann, Amy Apprill, Markus Bauer, Donna Berg-Lyons, Jason Betley, T. D. Bruns, J. Greg Caporaso, Noah Fierer, Louise Fraser, Jed A. Fuhrman, M. Gardes, Jack A. Gilbert, Niall Gormley, Greg Humphrey, James Huntley, Janet...
["[Amplification Protocol] Amplify samples in triplicate.", "[Amplification Protocol] Pool triplicate PCR reactions for each sample into a single volume (75 µL). Do not combine amplicons from different samples at this point.", "[Amplification Protocol] Run amplicons from each sample on an agarose gel.", "[Amplification...
40,922
ELISA for quantification of human C3 in serum or plasma.
6
dx.doi.org/10.17504/protocols.io.bj72krqe
https://www.protocols.io/view/elisa-for-quantification-of-human-c3-in-serum-or-bj72krqe
Angel Justiz-Vaillant, Belkis Ferrer-Cosme
TITLE: ELISA for quantification of human C3 in serum or plasma. AUTHORS: Angel Justiz-Vaillant, Belkis Ferrer-Cosme [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. An anti-human C3 coating antibody is adsorbed onto the microwells by incubation overnight at 4°C with carbonate-bicarbonate buffer. ?....
["An anti-human C3 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 C3 present in the serum or plasma binds to antibodies adsorbed into the microwells.", "The microplate is blocked with 3% non-fat milk-PBS buf...
101,063
12S rRNA-Gene Metabarcoding Library Prep: Dual-PCR Method
1
dx.doi.org/10.17504/protocols.io.n2bvjnn25gk5/v1
https://www.protocols.io/view/12s-rrna-gene-metabarcoding-library-prep-dual-pcr-dexf3fjn
rute.carvalho Carvalho, Colleen Kellogg, Matt Lemay
TITLE: 12S rRNA-Gene Metabarcoding Library Prep: Dual-PCR Method AUTHORS: rute.carvalho Carvalho, Colleen Kellogg, Matt Lemay [DESCRIPTION] This protocol is used for eDNA metabarcoding of the mitochondrial 12S rRNA gene (Miya et al 2015) using Pair-End Illumina Miseq Sequencing. As part of the Hakai Institute Ocean Ob...
["[Preparations] Ensure that the laboratory is appropriately configured and that staff has appropriate training. See \"Guidelines\" for more information. Pay attention to the separation of pre and post-PCR spaces and equipment.", "[Triplicate PCR Amplification (1st PCR)] Preparations\n\n \n\n Reagents:\n (Or equal)\n ...
83,165
Concentration and nucleic acid extraction of viruses from wastewater influent
4
null
https://www.protocols.io/view/concentration-and-nucleic-acid-extraction-of-virus-cvf5w3q6
Ari N Machtinger, Olivia S Hershey, William J Bradshaw, Michael R McLaren
TITLE: Concentration and nucleic acid extraction of viruses from wastewater influent AUTHORS: Ari N Machtinger, Olivia S Hershey, William J Bradshaw, Michael R McLaren [DESCRIPTION] In this protocol, 200 mL of raw influent wastewater is concentrated to a final volume of 400 uL using the Innovaprep Concentrating Pipet...
["[Part 1: Influent Handling, Dissociation, Centrifugation, Filtration] In the fume hood, add 400 uL of 10% Tween 20 stock solution each to the seven centrifuge tubes.", "[Part 1: Influent Handling, Dissociation, Centrifugation, Filtration] Prepare the negative control. Add 40 mL of PBS to two of the centrifuge tubes....
93,627
Practical Guide to Live Sampling of Livestock and Wildlife for Infectious Disease Surveillance
1
dx.doi.org/10.17504/protocols.io.rm7vzxro8gx1/v2
https://www.protocols.io/view/practical-guide-to-live-sampling-of-livestock-and-c7n3zmgn
Stefano Catalano
TITLE: Practical Guide to Live Sampling of Livestock and Wildlife for Infectious Disease Surveillance AUTHORS: Stefano Catalano [DESCRIPTION] Under- or misdiagnosed cases of disease caused by especially dangerous pathogens present public health and proliferation risks. Numerous studies have demonstrated that in low-re...
[]
54,317
Consistency in Identity Related Sequential Decisions
2
null
https://www.protocols.io/view/consistency-in-identity-related-sequential-decisio-bzamp2c6
Dikla Perez, Yael Steinhart, Amir Grinstein, Meike Morren
TITLE: Consistency in Identity Related Sequential Decisions AUTHORS: Dikla Perez, Yael Steinhart, Amir Grinstein, Meike Morren [DESCRIPTION] We conducted four lab and online experiments, and a field experiment to test our hypotheses and to rule out an alternative explanation. The design of each of the five experimen...
[]
97,050
Introduction to flux balance analysis (FBA)
0
null
https://www.protocols.io/view/introduction-to-flux-balance-analysis-fba-daz22f8e
Cailean Carter, Dipali Singh, Gemma Langridge
TITLE: Introduction to flux balance analysis (FBA) AUTHORS: Cailean Carter, Dipali Singh, Gemma Langridge [DESCRIPTION] Flux balance analysis (FBA) is a mathematical approach to finding an optimal net flow of mass through a metabolic network that follows a set of instructions defined by the user. This protocol covers ...
["[Background] Flux balance analysis (FBA) is a mathematical approach to finding an optimal net flow of mass through a metabolic network that follows a set of instructions defined by the user. This protocol covers the mathematical principles behind FBA and provides coding examples that can be followed using Python3. A ...
null
null
null
dx.doi.org/10.17504/protocols.io.cp8vrv
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.erabd2e
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <p>Components:<br /><br />2.25 M DTT, 40 mM KOAc, pH 6.0.</p> [STEPS] ?. ?. ?.
[]
41,181
manu 3
1
null
https://www.protocols.io/view/manu-3-bkf5ktq6
Monica Hassan
TITLE: manu 3 AUTHORS: Monica Hassan [STEPS] ?.
[]
32,624
Single molecule FISH
null
dx.doi.org/10.17504/protocols.io.bb4qiqvw
null
Thuc Nguyen, Emma Garren
TITLE: Single molecule FISH AUTHORS: Thuc Nguyen, Emma Garren [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Updated This protocol describes multiround hybrization of directly-conjugated FISH probes for single molecule RNA detection. Thin tissue sections (10-μm) are placed onto silanized coverslips...
["[Coverslip Preparation]\nClean coverslips (Thorlabs #CG15KH) with lens paper and 70% ethanol. With minimal handling of the coverslips (touch edges with clean gloves is ok), load them into a coverslip rack compatible with a plasma cleaning oven. We use a quartz coverslip rack. Placing coverslips in a clean glass conta...
83,013
Microscopy-based bead protein-protein interaction assay
1
dx.doi.org/10.17504/protocols.io.dm6gp3erpvzp/v1
https://www.protocols.io/view/microscopy-based-bead-protein-protein-interaction-cvbdw2i6
Elisabeth Holzer
TITLE: Microscopy-based bead protein-protein interaction assay AUTHORS: Elisabeth Holzer [DESCRIPTION] This protocol describes how to perform microscopy-based bead protein-protein interaction assay with GST- or mCherry-tagged proteins as baits and fluorescently-tagged proteins as preys. The protocol requires to have p...
["[Prepare bait-coated beads] Equilibrate 20 µL Glutathione Sepharose 4B or RFP-Trap Agarose beads with 200 µL SEC buffer", "[Prepare bait-coated beads] Incubate equilibrated beads with GST- or mCherry-tagged bait protein for a final concentration of 5 micromolar (µM) in SEC buffer for 60 min at 4 °C with gentle rotati...
28,245
MojoSort™ Selection Kits Column Protocol - 3
null
dx.doi.org/10.17504/protocols.io.7tvhnn6
null
Sam Li
TITLE: MojoSort™ Selection Kits Column Protocol - 3 AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>BioLegend MojoSort™ nanobeads work in commonly used separation columns, based on our internal research as well as validation by external testing by academic labs. This simple pro...
["Prepare cells from your tissue of interest or blood without lysing erythrocytes. Kits for human samples have been optimized for PBMCs, please prepare the cells using a suitable method.", "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) ...
97,425
RNA Slide Preparation Protocol (FFPE) for nanostring DSP - GeoMx - Human Ovary U54 Native Tissue v2
4
dx.doi.org/10.17504/protocols.io.bp2l624bdgqe/v2
https://www.protocols.io/view/rna-slide-preparation-protocol-ffpe-for-nanostring-dbdr2i56
Nicolas Martin, Tommy Tran
TITLE: RNA Slide Preparation Protocol (FFPE) for nanostring DSP - GeoMx - Human Ovary U54 Native Tissue v2 AUTHORS: Nicolas Martin, Tommy Tran [DESCRIPTION] This protocol is designed for RNA slide preparation for formalin-fixed tissue. [GUIDELINES] This is the default protocol for slide preparation by Nanostring. ...
["Prepare reagents\n\nPrepare the reagents using the dilution instructions (see Table 1).\nUse DEPC- treated water for all dilutions. The actual volume of\nreagents used in the protocol will vary – the volumes to prepare in Table 1 are suggestions.\n\nTable 1: Reagent prep for RNA slide preparation\n\n Reagent ...
38,666
Intravenous Jugular Catheterization for Rats
1
null
https://www.protocols.io/view/intravenous-jugular-catheterization-for-rats-bhzij74e
Sharona Sedighim, Lani Tieu, Olivier George
TITLE: Intravenous Jugular Catheterization for Rats AUTHORS: Sharona Sedighim, Lani Tieu, Olivier George [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol outlines the step-by-step setup and procedure of the pre-op, surgery, and post-op for intravenous jugular cathetarization for rats in...
["[Animal Preparation for Surgery]\nPlace rat in the knock-down chamber. Set isoflurane vaporizer dial on 5 to begin anesthetizing rat.", "[Animal Preparation for Surgery]\nOnce breathing is constant (about 2 breaths every 3 seconds) and deep, turn off isoflurane and take the animal out for preparation.", "[Animal Prep...
50,058
PEI/Laminin Coating
1
dx.doi.org/10.17504/protocols.io.bu5iny4e
https://www.protocols.io/view/pei-laminin-coating-bu5iny4e
Zoe
TITLE: PEI/Laminin Coating AUTHORS: Zoe [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is used to prepare plates for iPSC-derived neuron maturation. It is a part of the NGN2 cortical neuron differentiation protocol: coat plates in preparation for Day 7 dissociation and replating.</di...
["[PEI Coating]\nAt least one day before plating, make 2x borate buffer with 20x borate buffer (Thermo, cat. no. 28341) in water in a Falcon tube.\n4 mL\n36 mL\n50 mL", "[PEI Coating]\nDilute 5% PEI (poly-ethylenimine; stored at ) in 2x borate buffer to make 0.1% PEI.\n800 µl\n4 °C\n40 mL", "[PEI Coating]\nFilter-...
22,183
Euplotes crassus micronuclear enrichment by PFGE
null
dx.doi.org/10.17504/protocols.io.zwff7bn
null
Rachele Cesaroni, Angela Piersanti
TITLE: Euplotes crassus micronuclear enrichment by PFGE AUTHORS: Rachele Cesaroni, Angela Piersanti [STEPS] ?. Starved Euplotes crassus cells (treated overnight with ampicillin 100 mg/ml) were filtered and harvested by centrifugation at 400 rcf for 3 min. ?. As much as possible of the sea water was removed and cells w...
["Starved Euplotes crassus cells (treated overnight with ampicillin 100 mg/ml) were filtered and harvested by centrifugation at 400 rcf for 3 min.", "As much as possible of the sea water was removed and cells were resuspended in 1V of 1XTE buffer and mixed with 1V of 2% Certified Low-Melt Agarose (Bio-Rad) in 1XTE buf...
77,722
In situ CD79a detection in formalin-fixed, paraffin-embedded (FFPE) pig tissues
4
dx.doi.org/10.17504/protocols.io.x54v9dzq1g3e/v1
https://www.protocols.io/view/in-situ-cd79a-detection-in-formalin-fixed-paraffin-cp52vq8e
Jayne E Wiarda, Crystal Loving
TITLE: In situ CD79a detection in formalin-fixed, paraffin-embedded (FFPE) pig tissues AUTHORS: Jayne E Wiarda, Crystal Loving [DESCRIPTION] An immunohistochemistry (IHC) staining protocol for in situ identification of porcine CD79a [BEFORE_START] Starting specimens: Starting samples = FFPE tissues cut to 4 micron th...
["[Baking] Before starting the assay: \nPreheat a dry oven to 60℃ \nLoad slides for assay into vertical slide rack\n\nBaking\nBake slides 20 min 60℃\n\nWhile slides bake:\nPrepare 0.05% PBS-T (can store at RT up to 1 month)", "[Deparaffinizing &amp; Rehydrating] Immediately before deparaffinizing:\nAdd ~200 mL xylenes ...
62,752
Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson’s disease: Cell-Based in vitro Assays
4
dx.doi.org/10.17504/protocols.io.5jyl89648v2w/v1
https://www.protocols.io/view/regulation-of-mitophagy-by-the-nsl-complex-underli-b9h8r39w
Marc P.M. Soutar, Daniela Melandri, Benjamin O'Callaghan, Emily Annuario, Amy E. Monaghan, Paul J. Whiting, Helene Plun-Favreau
TITLE: Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson’s disease: Cell-Based in vitro Assays AUTHORS: Marc P.M. Soutar, Daniela Melandri, Benjamin O'Callaghan, Emily Annuario, Amy E. Monaghan, Paul J. Whiting, Helene Plun-Favreau [DESCRIPTION] Impaired mitophagy is a key causative pat...
["[Cell Culture and siRNA Transfection] Culture cells in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, 11995-0 65) supplemented with 10% heat-inactivated foetal bovine serum (FBS, Gibco) in a humidified chamber at 37 °C with 5% CO2.", "[Cell Culture and siRNA Transfection] For siRNA transfection, transfect cells using...
46,098
My new protocol
1
dx.doi.org/10.17504/protocols.io.bq9smz6e
https://www.protocols.io/view/my-new-protocol-bq9smz6e
Monica Hassan
TITLE: My new protocol AUTHORS: Monica Hassan [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">any</div></div> [STEPS] ?. Pressure: ?.
["Pressure:"]
62,525
OcuPrime - 100% Safe And Effective For Eyes!
3
dx.doi.org/10.17504/protocols.io.j8nlkkn6wl5r/v1
https://www.protocols.io/view/ocuprime-100-safe-and-effective-for-eyes-b9a5r2g6
OcuPrime
TITLE: OcuPrime - 100% Safe And Effective For Eyes! AUTHORS: OcuPrime [DESCRIPTION] OcuPrime [STEPS]
[]
66,100
Monitoring the point spread function for quality control of confocal microscopes
1
dx.doi.org/10.17504/protocols.io.bp2l61ww1vqe/v1
https://www.protocols.io/view/monitoring-the-point-spread-function-for-quality-ccsuswew
Glyn Nelson, Ioannis Alexopoulos, Maria Azevedo, Fabio Barachati, Yury Belyaev, Mariana T Carvalho, Yann Cesbron, Aurelien Dauphin, Alexander D Corbett, Ian M Dobbie, Laurent Gelman, Nadia Halidi, Xiang Hao, Hella Hartmann, Rainer Heintzmann, Peter Hemmerich, Marcel Kirchner, Judith Lacoste, Penghuan Liu, Laure Planta...
TITLE: Monitoring the point spread function for quality control of confocal microscopes AUTHORS: Glyn Nelson, Ioannis Alexopoulos, Maria Azevedo, Fabio Barachati, Yury Belyaev, Mariana T Carvalho, Yann Cesbron, Aurelien Dauphin, Alexander D Corbett, Ian M Dobbie, Laurent Gelman, Nadia Halidi, Xiang Hao, Hella Hartmann...
["[Bead Slide Preparation] The bead slide preparation described below contains steps previously published in the following protocols:\n \nas well as another:", "[Bead Slide Preparation] Bead dilution", "[Bead Slide Preparation] Bead slide mounting", "[Bead Slide Preparation] The bead slides can be stored at room temper...
65,392
Pure Calms CBD Gummies – Reviews &amp; Price 2022
1
dx.doi.org/10.17504/protocols.io.kqdg3pn71l25/v1
https://www.protocols.io/view/pure-calms-cbd-gummies-reviews-amp-price-2022-cb4qsqvw
purecalmot
TITLE: Pure Calms CBD Gummies – Reviews &amp; Price 2022 AUTHORS: purecalmot [DESCRIPTION] Pure Calms CBD Gummies is the item that is ideal to accomplish calming, cleaning, and mitigating properties for muscle joints, nerves, nails, hair, skin, joints, and some more. This item is particularly alright for outside uti...
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null
null
null
dx.doi.org/10.17504/protocols.io.cicuav
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
50,425
Plankton DNA extraction from Sterivex filter units
4
dx.doi.org/10.17504/protocols.io.bvgzn3x6
https://www.protocols.io/view/plankton-dna-extraction-from-sterivex-filter-units-bvgzn3x6
Marine Vautier, Cécile Chardon, Camilla Capelli, Rainer Kurmayer, Nico Salmaso, Isabelle Domaizon
TITLE: Plankton DNA extraction from Sterivex filter units AUTHORS: Marine Vautier, Cécile Chardon, Camilla Capelli, Rainer Kurmayer, Nico Salmaso, Isabelle Domaizon [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The objective of this protocol is to provide a reliable and replicable method for the...
["Take the Sterivex™ filter unit (previously stored at after filtration). Remove the Inlet Cap (Figure 1) and add of Solution ST1B using a pipette tip. Note: Insert the tip completely into the inlet so that the pipette tip is visible inside the unit just above the membrane. Note: Solution ST1A must be added to bottl...
99,621
Protocol for development and temporal in vivo imaging of RVO mouse models
0
dx.doi.org/10.17504/protocols.io.ewov19nrplr2/v1
https://www.protocols.io/view/protocol-for-development-and-temporal-in-vivo-imag-ddid24a6
Xu Xiaowei
TITLE: Protocol for development and temporal in vivo imaging of RVO mouse models AUTHORS: Xu Xiaowei [DESCRIPTION] This is an protocol for development and temporal in vivo imaging of RVO mouse models. [STEPS] SECTION: Animals and Anesthesia 1. Male C57BL/6J mice, aged 6-8 weeks and weighing 19-21g, were purchased fr...
["[Animals and Anesthesia] Male C57BL/6J mice, aged 6-8 weeks and weighing 19-21g, were purchased from DOSSY EXPERIMENTAL ANIMALS CO. LTD, Chengdu, China. The mice were adaptively raised in an environment with a 12h/12h light/dark cycle, constant room temperature of 26°C, and adequate water and food for one week before...
18,106
New version of protocol 2 images pokipoki
null
dx.doi.org/10.17504/protocols.io.vw2e7ge
null
Darja Darja
TITLE: New version of protocol 2 images pokipoki AUTHORS: Darja Darja [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Between May and June 1993, Slovenia competed in the qualifiers for the EuroBasket 1993, where the team won all seven games and therefore qualified for its first EuroBasket. At the ...
["[1. ball ]\nBetween May and June 1993, Slovenia competed in the qualifiers for the EuroBasket 1993, where the team won all seven games and therefore qualified for its first EuroBasket. At the main tournament, held in Germany, Slovenia finished in 14th place out of 16 teams with one win and two defeats. Between Ma...
53,981
Thawing primary leukemia cells
4
dx.doi.org/10.17504/protocols.io.byx5pxq6
https://www.protocols.io/view/thawing-primary-leukemia-cells-byx5pxq6
Kathrin Bernt
TITLE: Thawing primary leukemia cells AUTHORS: Kathrin Bernt [DESCRIPTION] This protocol is used to thaw primary cells. Key points are the addition of DNAse, which helps to preserve viability. [STEPS] 1. Transfer the contents to a 15ml conical tube. 2. Add DNase. 1/10 of the volume of liquid in the tube (100µl/1ml o...
["Transfer the contents to a 15ml conical tube.", "Add DNase. 1/10 of the volume of liquid in the tube (100µl/1ml of sample). Do not pipette. Gently shake the sample to mix in the DNase.", "Incubate sample in 37˚C water bath for 90 seconds.", "Add media. 10ml, Drop for drop or very slowly while again gently mixing the ...
98,951
Immunofluorescence Staining in Mouse Brain Tissue Sections
0
dx.doi.org/10.17504/protocols.io.14egn6zopl5d/v1
https://www.protocols.io/view/immunofluorescence-staining-in-mouse-brain-tissue-dcvf2w3n
madalynn.erb Erb
TITLE: Immunofluorescence Staining in Mouse Brain Tissue Sections AUTHORS: madalynn.erb Erb [DESCRIPTION] This protocol details the immunofluorescence staining in mouse brain tissue sections. [STEPS] SECTION: Day 1 1. Staining of 35μm free-floating mouse brain sections is performed in glass staining dishes (Pyrex 367...
["[Day 1] Staining of 35μm free-floating mouse brain sections is performed in glass staining dishes (Pyrex 36754-60) using 8-section staining nets (Ted Pella 36154-64). dx.doi.org/10.17504/protocols.io.5jyl8pzk9g2w/v1", "[Day 1] The sections can be transferred between wells using a paint brush.", "[Day 1] Volume of sol...
40,375
MPAPASS - Creating an MPAPASS database
1
dx.doi.org/10.17504/protocols.io.bjnxkmfn
https://www.protocols.io/view/mpapass-creating-an-mpapass-database-bjnxkmfn
Joshua Welsh, Sean Cook, Jennifer Jones
TITLE: MPAPASS - Creating an MPAPASS database AUTHORS: Joshua Welsh, Sean Cook, Jennifer Jones [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is collection contains the protocols required for each step in the mpapass software pipeline for performing stitched multiplex analysis. This is one of ...
["[Open New Dataset Window]\nOpen the MPAPASS software and navigate to the Menu tab in the upper left-hand corner. Under the Menu tab, choose the New Dataset option and a new window will pop-up as shown below:", "[Select CSV Directory]\nTo construct the database, follow the steps outlined on the right side of the windo...
74,334
QIAGEN DNeasy PowerMax Soil Kit
4
dx.doi.org/10.17504/protocols.io.6qpvr4ke3gmk/v1
https://www.protocols.io/view/qiagen-dneasy-powermax-soil-kit-ckt6uwre
QIAGEN
TITLE: QIAGEN DNeasy PowerMax Soil Kit AUTHORS: QIAGEN [DESCRIPTION] For the isolation of microbial DNA from large quantities of soil - great for samples with low microbial load The DNeasy PowerMax Soil Kit comprises a novel and proprietary method for isolating genomic DNA from environmental samples using Inhibitor R...
["[Sample preparation &amp; cell lysis] ADD 15 mL of PowerBead Solution to a PowerMax Bead Pro Tube\n\nADD up to 10 g of soil sample to the PowerMax Bead Pro Tube containing PowerBead Solution\n\nVORTEX vigorously for 1 min", "[Sample preparation &amp; cell lysis] ADD 1.2 mL of Solution C1 to the PowerMax Bead Pro Tube...
62,244
Forti Prime Italy : How To Use It In 2022! {UPDATED NEWS}
1
dx.doi.org/10.17504/protocols.io.kqdg3p747l25/v1
https://www.protocols.io/view/forti-prime-italy-how-to-use-it-in-2022-updated-ne-b82cryaw
hooksrobs
TITLE: Forti Prime Italy : How To Use It In 2022! {UPDATED NEWS} AUTHORS: hooksrobs [DESCRIPTION] Forti Prime Reviews: Forti Prime is an all-natural immune booster formula that includes a powerful combination of the world’s best immune-boosting ingredients. 100% safe to use a supplement, Check out its dosage, benefi...
["[Forti Prime]"]
null
null
null
dx.doi.org/10.17504/protocols.io.dh338m
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Combine<br />2mls glacial acetic acid<br />1L distilled water [STEPS]
[]
79,780
Gcase co-immunoprecipitation
1
dx.doi.org/10.17504/protocols.io.kxygx9xozg8j/v1
https://www.protocols.io/view/gcase-co-immunoprecipitation-cr6cv9aw
michela.deleidi, Federico Bertoli
TITLE: Gcase co-immunoprecipitation AUTHORS: michela.deleidi, Federico Bertoli [DESCRIPTION] We developed this protocol to identify protein-protein interactions between the enzyme glucocerebrosidase (GCase) and other proteins in human iPSC-derived Neural Precursor Cells. [STEPS] SECTION: Gcase co-immunoprecipitatio...
["[Gcase co-immunoprecipitation] Wash cells 1X with phosphate-buffered saline (PBS, Sigma‒Aldrich) and detach using Accutase.", "[Gcase co-immunoprecipitation] Pellet the cell suspension at 280 rcf, 5 min, 23 °C.", "[Gcase co-immunoprecipitation] Lyse the pellets in IP/lysis buffer (Thermo Fisher, #87787) supplemented ...
81,154
5&#39; RACE for RNA fragments with 5&#39; phosphate
4
dx.doi.org/10.17504/protocols.io.261ge3mzwl47/v1
https://www.protocols.io/view/5-39-race-for-rna-fragments-with-5-39-phosphate-cthawj2e
Marta.Gaglia
TITLE: 5&#39; RACE for RNA fragments with 5&#39; phosphate AUTHORS: Marta.Gaglia [DESCRIPTION] This protocol will allow identification of fragments of RNA that have a 5' phosphate. It relies on ligation of an RNA adapter to the 5' end of the fragment and amplification with primers that bind the adapter and gene-specif...
["RNA purification:\nExtract RNA and elute/resuspend in 50 µl H2O", "DNase treatment: \n\nTo each 50 µl sample, add 10 µl of DNase reaction mix containing:\n6 µl DNase buffer\n1.2 µl Turbo DNase\n1.5 µl RNasin\n1.3 µl H2O\n(total volume = 60 µl)\n\nIncubate for 15 min at 37°C (waterbath or heatblock).", "RNA extraction...
28,181
Aortic Banding in Mice
null
dx.doi.org/10.17504/protocols.io.7rvhm66
null
Dale Abel
TITLE: Aortic Banding in Mice AUTHORS: Dale Abel [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This protocol describes the procedure used by the DiaComp for inducing pressure overload hypertrophy in mice.</div><div c...
["Mice are anesthetized with 1% isoflurane delivered via nose cone. A topical depilatory agent is applied to the neck and chest and the area is cleaned with betadine and alcohol. Mice are placed supine and temperature maintained at 37ºC with a heating pad. A horizontal skin incision ~ 0.5 - 1.0 cm in length is made at ...
68,593
Targeted isolation of circular extrachromosomal DNA by CRISPR-CATCH
4
dx.doi.org/10.17504/protocols.io.ewov1ne8ygr2/v1
https://www.protocols.io/view/targeted-isolation-of-circular-extrachromosomal-dn-ce8rthv6
King L Hung, Howard Y. Chang
TITLE: Targeted isolation of circular extrachromosomal DNA by CRISPR-CATCH AUTHORS: King L Hung, Howard Y. Chang [DESCRIPTION] This protocol enables targeted isolation of circular extrachromosomal DNA using CRISPR-Cas9-mediated linearization followed by pulsed field gel electrophoresis. Isolated DNA products can be su...
["[Embedding cells in agarose plugs] Melt 1% Certified Low Melt Agarose solution in PBS, place in 45C water bath.", "[Embedding cells in agarose plugs] Pellet 1 million cells per agarose plug at 300g for 5 minutes. (for example, spin down 10 million cells to make 10 agarose plugs)", "[Embedding cells in agarose plugs] ...
54,433
Adult mouse kidney dissociation (on ice)
1
dx.doi.org/10.17504/protocols.io.bzd9p296
https://www.protocols.io/view/adult-mouse-kidney-dissociation-on-ice-bzd9p296
Andrew Potter
TITLE: Adult mouse kidney dissociation (on ice) AUTHORS: Andrew Potter [DESCRIPTION] Protocol for adult (8-10 week) mouse kidney dissociation performed on ice to reduce artifact gene expression. The protocol is based on our protocol published in Development for P1 mouse kidney, however this protocol includes two layer...
["[Isolate kidney] Coarsely mince tissue in PBS. \n\n2 min", "[Layer 1] Weigh out 25 mg coarsely minced tissue for each set of kidneys (remove PBS before weighing).\n\n25 5", "[Layer 1] Incubate digest mix for 10 min on ice with trituration and shaking. Triturate 15 strokes using 1 mL pipet set to 600 µL every 2 min; s...
25,002
Plasmid used for transfection trails of Hematodinium
null
dx.doi.org/10.17504/protocols.io.4nigvce
null
Imen Lassadi
TITLE: Plasmid used for transfection trails of Hematodinium AUTHORS: Imen Lassadi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Attempts to transfect</span><span style = "font-style:italic;"> Hematodinium </span><span>was performed using a plasmid that will express eGFP under the control o...
[]
69,810
Testing the effect of paraquat on C. elegans behaviour when on Keio E. coli mutants (6-well plates)
1
dx.doi.org/10.17504/protocols.io.6qpvr4e6zgmk/v1
https://www.protocols.io/view/testing-the-effect-of-paraquat-on-c-elegans-behavi-cgesttee
Saul Moore
TITLE: Testing the effect of paraquat on C. elegans behaviour when on Keio E. coli mutants (6-well plates) AUTHORS: Saul Moore [DESCRIPTION] Protocol for screening candidate behaviour-modifying E. coli BW25113 single-gene deletion mutants from the 'Keio Collection', to investigate their differential effects on Caeno...
["[Preparing NGM agar + pouring plates] Prior to screening, prepare the materials needed for screening C. elegans on selected Keio E. coli mutants:\n\n- 6-well plates (aka. 'imaging plates')\n- 15 mL Falcon tubes\n- 50 mL Erlenmeyer flasks\n- 90 mm Petri plates (aka. 'maintenance plates')\n- 150 mm Petri plates (aka. '...
24,353
Step-by-step protocol for high resolution respirometry for human heart homogenates
null
dx.doi.org/10.17504/protocols.io.3z9gp96
null
Adéla Krajčová, Tomáš Urban, Petr Waldauf, František Duška
TITLE: Step-by-step protocol for high resolution respirometry for human heart homogenates AUTHORS: Adéla Krajčová, Tomáš Urban, Petr Waldauf, František Duška [STEPS] ?. [Step-by-step protocol for high resolution respirometry for human heart homogenates] Firstly, wash properly all tubes, scissors, forceps, teflon pestl...
["[Step-by-step protocol for high resolution respirometry for human heart homogenates]\nFirstly, wash properly all tubes, scissors, forceps, teflon pestles and glassware (including Dounce Tissue homogenizer and glass pestles) with 70% ethanol and tap water and keep them on ice to cool to 0°C. Immediatelly, before use, ...
45,555
2. Gel run and transfer -Tricine
4
null
https://www.protocols.io/view/2-gel-run-and-transfer-tricine-bqqtmvwn
Elizabeth Fozo
TITLE: 2. Gel run and transfer -Tricine AUTHORS: Elizabeth Fozo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Western Blot</span></div></div> [STEPS] ?. [Buffers:] Tricine SDSRunningBuffer- dilute from 10 X to 1X Composition 10X/L100...
["[Buffers:]\nTricine SDSRunningBuffer- dilute from 10 X to 1X Composition 10X/L100 mM Tris base (121 g)100 mM Tricine (179 g)0.1% SDS (10g)pH 8.3The pH of the 1X solution is 8.3. Do not use acid or base to adjust pH (Novex, 2003)St...
50,781
Large scale screening of SARS-CoV-2 variants of worldwide concern by RT-qPCR
1
dx.doi.org/10.17504/protocols.io.bvt5n6q6
https://www.protocols.io/view/large-scale-screening-of-sars-cov-2-variants-of-wo-bvt5n6q6
Katja Spiess , Ellinor Marving, Alonzo Alfaro-Núñez, Vithiagaran Gunalan, Sofie Holdflod Nielsen, Michelle G. P. Jørgensen, Anna S. Fomsgaard, Søren M. Karst, Shila Mortensen, Ria Lassaunière, Jannik Fonager, Morten Rasmussen, Maiken Worsøe Rosenstierne, Charlotta Polacek Strandh, Anne-Marie Vangsted, Claus Nielsen, A...
TITLE: Large scale screening of SARS-CoV-2 variants of worldwide concern by RT-qPCR AUTHORS: Katja Spiess , Ellinor Marving, Alonzo Alfaro-Núñez, Vithiagaran Gunalan, Sofie Holdflod Nielsen, Michelle G. P. Jørgensen, Anna S. Fomsgaard, Søren M. Karst, Shila Mortensen, Ria Lassaunière, Jannik Fonager, Morten Rasmussen,...
["Protocol for 96 and 384 well PCR formatPrepare three different mastermixes. Make sure to vortex all reagents before use.Mastermix 1, for detection of H69/V70 and N501Y, contains the following: Mastermix 2, for detection of E484K, contains the following: Mastermix 3, for detection of L452R, contains the following: Add...
80,628
Neural differentiation on EM grids - iNeurons sample preparation for cryo-ET and CLEM
4
null
https://www.protocols.io/view/neural-differentiation-on-em-grids-ineurons-sample-csyuwfww
Cristina Capitanio, Victoria Trinkaus, Melissa Hoyer
TITLE: Neural differentiation on EM grids - iNeurons sample preparation for cryo-ET and CLEM AUTHORS: Cristina Capitanio, Victoria Trinkaus, Melissa Hoyer [DESCRIPTION] This is a protocol for differentiating AAVS1-TRE3G-NGN2 iPSCs and hESCs to iNeurons directly on EM grids for cryo-ET and cryo-CLEM. The protocol co...
["[Starting the neural differentiation (Day 0-Day6)] We based our neural differentiation protocol, for both iPSCs and hESCs AAVS1-TRE3G-NGN2 cells, on: \n \nWe also refer to that protocol for a complete list of media and reagents. \n\nFollow the neural differentiation protocol until cell splitting on Day 6;\nOne well o...
60,358
Highly Parallel Droplet Sample Preparation for Single Cell Proteomics
1
dx.doi.org/10.17504/protocols.io.b67erhje
https://www.protocols.io/view/highly-parallel-droplet-sample-preparation-for-sin-b67erhje
Andrew Leduc, Richard Huffman, Joshua Cantlon, Saad Khan, Nikolai Slavov
TITLE: Highly Parallel Droplet Sample Preparation for Single Cell Proteomics AUTHORS: Andrew Leduc, Richard Huffman, Joshua Cantlon, Saad Khan, Nikolai Slavov [DESCRIPTION] Protocol for preparing single cells for mass-spec analysis by nPOP as described by Leduc et al., 2021, 2022 DOI: 10.1101/2021.04.24.441211. n...
["[Carrier and Reference Preparation (***Only if Carrier is used***)] Prepare cell pellets of at least 500,000 cells for all relevant cell types . Add 100% DMSO to cells to a cellular concentration of 6000 cells/ul. Incubate cells in DMSO for 20 minutes to lyse cells. Add mass spectrometry grade water to bring solution...
36,056
SARS-CoV-2 Sequencing on Illumina MiSeq Using ARTIC Protocol: Part 2 - Illumina DNA Flex Protocol
null
dx.doi.org/10.17504/protocols.io.bffyjjpw
https://www.protocols.io/view/sars-cov-2-sequencing-on-illumina-miseq-using-arti-bffyjjpw
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 Flex Protocol AUTHORS: Joel Sevinsky, Arian Nassiri, Heather Blankenship, Erin Young, Kevin Libuit, Kelly Oakeson, Lauren Turner, StaPH-B Consortium [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protoco...
["[DNA Flex - Dilution Plate Preparation]\nDilution Plate Preparation Date/Initials:_________________Prior to starting your DNA Flex library prep, samples should be diluted into a 96 well plate as described below. Ideally, you would like each sample to be diluted such that the final volulme of sample contains ...
null
null
null
dx.doi.org/10.17504/protocols.io.dpf5jm
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] The modified dilution assay aims to partition phytoplankton mortality into virus- versus grazing-induced fractions and has previously been applied to several different environments to determine viral lysis rates of natural phytoplankton. The method involves creating a gradient o...
[]
62,496
Caltech-SCFA-methods fecal
1
dx.doi.org/10.17504/protocols.io.bp2l61rrkvqe/v1
https://www.protocols.io/view/caltech-scfa-methods-fecal-b898rz9w
rabdelha
TITLE: Caltech-SCFA-methods fecal AUTHORS: rabdelha [DESCRIPTION] This protocol details the Caltech-SCFA-methods for measurement of short-chain fatty acids in mouse fecal samples. [STEPS] SECTION: Sample preparation 2. Briefly add, ice-cold extraction solvent (1:1 v/v ACN/water) to fecal sample at a ratio of 2 µL:...
["[Sample preparation] Briefly add, ice-cold extraction solvent (1:1 v/v ACN/water) to fecal sample at a ratio of 2 µL: 1 mg sample and internal standard mix to a final concentration of 100 micromolar (µM) and subject to vortex mixing for 3 min at Room temperature and sonicate for 15 min.", "[Sample preparation] Centri...
null
null
null
dx.doi.org/10.17504/protocols.io.jsycnfw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Secreted protein, acidic and rich in cysteine (SPARC) is differentially associated with cell proliferation and extracellular matrix (ECM) assembly. We show here the effect of exogenous SPARC inhibition/induction on ECM and mitochondrial proteins expression and on the differen...
[]
48,981
How to increase the number of simultaneous users allowed in an R Shiny App
5
null
https://www.protocols.io/view/how-to-increase-the-number-of-simultaneous-users-a-bt3vnqn6
Sonia García-Ruiz
TITLE: How to increase the number of simultaneous users allowed in an R Shiny App AUTHORS: Sonia García-Ruiz [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">There are multiple ways of hosting a Shiny App in production. When it is required to do so over a specific URL domain, RStudio Server combined ...
["Install ShinyProxy.ShinyProxy is a java-based open-source tool to deploy Shiny apps in a production or enterprise context https://www.shinyproxy.io/.To install it on CentOS 7:sudo yum install shinyproxy_2.3.0_x86_64.rpm", "Configure ShinyProxy.Before executing any docker image, ShinyProxy first needs to be able to co...
76,263
Implant Surgery: Chronic recoverable Neuropixels in mice
1
dx.doi.org/10.17504/protocols.io.yxmvmnn2bg3p/v5
https://www.protocols.io/view/implant-surgery-chronic-recoverable-neuropixels-in-cnqfvdtn
Emily A Aery Jones
TITLE: Implant Surgery: Chronic recoverable Neuropixels in mice AUTHORS: Emily A Aery Jones [DESCRIPTION] This protocol collection explains how to build a low-cost, lightweight system to implant 1 Neuropixels 1.0 probe or 2 Neuropixels 2.0 probes into mice, record during freely moving behavior, then recover the probes...
["[Prepare surgical tools and field] Sterilize tips of metal instruments, ground screw, and headbar in autoclave 25 min or hot bead sterilizer 5 s . Disinfect cotton swabs, toothpick, and a weigh boat under UV light 30 min .", "[Prepare surgical tools and field] Disinfect surgical field with 10% bleach. Lay absorbent ...
20,627
Case - Intraperitoneal Glucose Tolerance Test
null
dx.doi.org/10.17504/protocols.io.ydtfs6n
null
Henri Brunengraber
TITLE: Case - Intraperitoneal Glucose Tolerance Test AUTHORS: Henri Brunengraber [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This is the standard protocol for most routine glucose tolerance testing. It is performe...
["Fast mice overnight or for 6 hours. Remove mouse from cage and put into a clean cage with water and no food (5:00PM). Next day begin GTT by 9:00AM.", "Weigh each mouse and record weight.", "Insert glucose strip into glucometer and check that the code matches for the strip being used.", "Take fasting blood glucose by ...
32,997
Testing repetitive tDCS on daily smoking behaviour: A placebo controlled EMA study
null
dx.doi.org/10.17504/protocols.io.bcgdits6
https://www.protocols.io/view/testing-repetitive-tdcs-on-daily-smoking-behaviour-bcgdits6
Ilse Verveer, Danielle Remmerswaal, Joran Jongerling, Frederik M. van der Veen, Ingmar H. A. Franken
TITLE: Testing repetitive tDCS on daily smoking behaviour: A placebo controlled EMA study AUTHORS: Ilse Verveer, Danielle Remmerswaal, Joran Jongerling, Frederik M. van der Veen, Ingmar H. A. Franken [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>The aim of the study was to explore the effect...
[]
81,613
TS Procure 812 - primary fixation formalin/wet tissue (TM - 013)
4
dx.doi.org/10.17504/protocols.io.bp2l61qbdvqe/v2
https://www.protocols.io/view/ts-procure-812-primary-fixation-formalin-wet-tissu-ctxmwpk6
sandra.crameri
TITLE: TS Procure 812 - primary fixation formalin/wet tissue (TM - 013) AUTHORS: sandra.crameri [DESCRIPTION] The method is used to process formalin fixed histological tissues to Procure 812 resin blocks. [GUIDELINES] Sample is not optimal for EM, best to dissect and use only the outer 2mm of tissue mass. Sample is ...
["[HEADER] SAN:\n\n\n\n\nSPEC No:\n\n\n\n\nOPERATOR & STEPS:\n\n\n\n\nOPERATOR & STEPS:", "[Primary fixation] Formalin for undefined extended period of time at Room temperature", "[Conventional] 2.5 % volume in 0.1 Molarity (M) Sorenson's Phosphate buffer (pH 7.2, 300mosmol/kg) for at least 40 min or overnight", "[C...
34,600
DNA Fragmentation &amp; Library Construction
2
dx.doi.org/10.17504/protocols.io.bp2l6nr4zgqe/v1
https://www.protocols.io/view/dna-fragmentation-amp-library-construction-bd2gi8bw
Marina McCowin
TITLE: DNA Fragmentation &amp; Library Construction AUTHORS: Marina McCowin [DESCRIPTION] The KAPA HyperPlus Kit provides a versatile, streamlined DNA fragmentation and library construction protocol for the rapid preparation of libraries for Illumina sequencing. The one-tube chemistry and protocol improves the efficie...
[]
108,556
Chemical fixation of Solarion arianae for transmission electron microscopy
0
dx.doi.org/10.17504/protocols.io.kxygxyd5zl8j/v1
https://www.protocols.io/view/chemical-fixation-of-solarion-arianae-for-transmis-dm9k494w
Marek Valt, Pavla Hrubá
TITLE: Chemical fixation of Solarion arianae for transmission electron microscopy AUTHORS: Marek Valt, Pavla Hrubá [DESCRIPTION] This is an optimized version of the protocol for standard chemical fixation for transmission electron microscopy that was used for the fixation of the culture of Solarion arianae. [STEPS] ...
["[Cell harvest] Centrifuge a well-grown culture at 1250 g at 4 °C.", "[Chemical cell fixation] Immerse pellet with 2,5% glutaraldehyde fixative in 0,1M cacodylate buffer solution for 1h. Work on ice.", "[Cell harvest] Discard supernatant and collect pellet.", "[Chemical cell fixation] Wash with 0,1M cacodylate buffer,...
66,881
Fluorescence size exclusion chromatography (FSEC) from ATP13A2 microsomes
6
dx.doi.org/10.17504/protocols.io.81wgb6bpolpk/v1
https://www.protocols.io/view/fluorescence-size-exclusion-chromatography-fsec-fr-cdi9s4h6
Sue Sim, eunyong_park
TITLE: Fluorescence size exclusion chromatography (FSEC) from ATP13A2 microsomes AUTHORS: Sue Sim, eunyong_park [DESCRIPTION] Using fluorescence size exclusion chromatography (FSEC) to analyze ATP13A2 expression in microsomes [STEPS] 1. Thaw 100 μg microsomes on ice 2. Resuspend microsomes in Lysis Buffer and fina...
["Thaw 100 μg microsomes on ice", "Resuspend microsomes in Lysis Buffer and final volume 1% DDM/0.2% CHS (1X pellet, 3X Lysis Buffer, 1X 5% DDM/1% CHS) at 4 °C", "DDM: n-dodecyl-β-D-maltopyranoside (Anatrace)", "CHS: cholesteryl hemisuccinate (Anatrace)", "Solubilize by rotating end-over-end for 2 h at 4C", "Clarify ly...
36,591
Phosphate Buffered Saline (PBS)
null
dx.doi.org/10.17504/protocols.io.bfypjpvn
https://www.protocols.io/view/phosphate-buffered-saline-pbs-bfypjpvn
Neilier Junior
TITLE: Phosphate Buffered Saline (PBS) AUTHORS: Neilier Junior [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A buffer solution has the function of resisting changes in pH even when adding powerful acids or bases. However, in the physiological environment the buffered system also provides cofactors...
["[Phosphate Buffered Saline (PBS)]\nPrepare by dissolving , and in .\n150 mM NaCl10 mM Potassium Phosphate buffer\n[PBS]\n[NaCl]\n[K2HPO4 • 3H2O]\n[KH2PO4]\n[distilled water]", "[Phosphate Buffered Saline (PBS)]\nAdjust the pH before use.", "[Phosphate Buffered Saline (PBS)]\nA variation of PBS can also be prepared...
86,540
NucBarcoder - a bioinformatic pipeline to characterise the genetic basis of plant species differences
5
dx.doi.org/10.17504/protocols.io.5qpvo33rzv4o/v1
https://www.protocols.io/view/nucbarcoder-a-bioinformatic-pipeline-to-characteri-cyrkxv4w
Wu Huang, Alex Twyford, Peter Hollingsworth
TITLE: NucBarcoder - a bioinformatic pipeline to characterise the genetic basis of plant species differences AUTHORS: Wu Huang, Alex Twyford, Peter Hollingsworth [DESCRIPTION] There are now a significant number of studies that have sequenced multiple loci from the nuclear genomes of plants and these are available in p...
["[Overview of the data management and the structure of NucBarcoder]", "[Data cleaning and filtering] Data cleaning and filtering were performed at both the individual and locus levels. \nDifferent clean-up and filtering tools were used for aligned sequence formats and SNP matrices. VCFtools (0.1.17) (Danecek et al., 2...
null
null
null
dx.doi.org/10.17504/protocols.io.e2tbgen
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The cells targeted by the Nanobeads are either selected or depleted by incubating your sample with the directly conjugated magnetic particles. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the targeted cells, downstream ...
[]
54,914
Starlet Sea Anemone (Nematostella vectensis) 2021 Environmental Summary, Reptile &amp; Aquatics, Stowers Institute for Medical Research
3
dx.doi.org/10.17504/protocols.io.bzvap62e
https://www.protocols.io/view/starlet-sea-anemone-nematostella-vectensis-2021-en-bzvap62e
Shane C. Miller, Diana P Baumann, M. Shane Merryman
TITLE: Starlet Sea Anemone (Nematostella vectensis) 2021 Environmental Summary, Reptile &amp; Aquatics, Stowers Institute for Medical Research AUTHORS: Shane C. Miller, Diana P Baumann, M. Shane Merryman [DESCRIPTION] The starlet sea anemone (Nematostella vectensis) is an emerging model organism, and we have maintai...
[]
52,731
ADP-Glo kinase assay
4
dx.doi.org/10.17504/protocols.io.bxq3pmyn
https://www.protocols.io/view/adp-glo-kinase-assay-bxq3pmyn
Chunmei Chang
TITLE: ADP-Glo kinase assay AUTHORS: Chunmei Chang [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">An assay for measuring ULK1 kinase activity</div></div> [STEPS] ?. Make a mixture of 100 nM purified wild-type or kinase dead ULK1 complex with 5 µM ULKtide in the presence or absence of 100 µM ATP. ...
["Make a mixture of 100 nM purified wild-type or kinase dead ULK1 complex with 5 µM ULKtide in the presence or absence of 100 µM ATP. The total volume is 50 µL.", "Add 50 µL ATP-depletion reagent to terminate the reaction and deplete the unconsumed ATP.", "Incubate at room temperature for 40 min.", "Add 10 µL of kinase...
null
null
null
dx.doi.org/10.17504/protocols.io.ftjbnkn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Yeast is a well-characterized genome due to its small size and historical significance in genetics.  The website http://yeastgenome.org/ is a dedicated resource for yeast genomics.</p> <p> </p> <p>For this exercise, I want you to create a "Makefile" that will execute this ent...
[]
78,441
Protein Transfer using Bio-rad TransBlot Turbo with Turbo RTA Transfer Kit
4
null
https://www.protocols.io/view/protein-transfer-using-bio-rad-transblot-turbo-wit-cquhvwt6
Lynn Doran, Steven J Burgess
TITLE: Protein Transfer using Bio-rad TransBlot Turbo with Turbo RTA Transfer Kit AUTHORS: Lynn Doran, Steven J Burgess [DESCRIPTION] A protocol for transfer of proteins from acrylamide gels onto nitrocellulose membrane for immunoblot analysis. This protocol is based on the assumption that TGX pre-cast gels have bee...
["Place the gel in the middle of the membrane and roll to remove air-bubbles.", "Place the top stack on top of the gel, gently roll", "Close the cassette lid, taking care not to disturb the gel", "Place the cassette in the Trans-Blot Turbo and follow the instructions on the machine (Fast protocol TGX gel - 3 minutes)...
null
null
null
dx.doi.org/10.17504/protocols.io.i35cgq6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
84,802
LRRK1 expression and purification
4
dx.doi.org/10.17504/protocols.io.rm7vzx3b5gx1/v1
https://www.protocols.click/view/lrrk1-expression-and-purification-cw3axgie
Yu Xuan Lin, janice reimer
TITLE: LRRK1 expression and purification AUTHORS: Yu Xuan Lin, janice reimer [DESCRIPTION] Protein purification protocol for full-length LRRK1 as done by Leschziner and Reck-Peterson Labs. Original protocol by Janice M. Reimer and Yu Xuan Lin. [STEPS] SECTION: His6-Z-TEV-LRRK1 Purification 1. N-terminally tagged Hi...
["[His6-Z-TEV-LRRK1 Purification] N-terminally tagged His6-Z-TEV-LRRK1(FL) was expressed in Sf9 insect cells. Insect cells were infected with baculovirus and grown at 27°C for 3 days.", "[His6-Z-TEV-LRRK1 Purification] Cells were harvested and then resuspended in lysis buffer (50mM HEPES pH 7.4, 500 mM NaCl, 20 mM imid...
24,779
96-well Plate Growth Curve Setup
null
dx.doi.org/10.17504/protocols.io.4fjgtkn
null
Norman van Rhijn, Panagiotis Papastamoulis, Takanori Furukawa, Magnus Rattray, Elaine Bignell, Mike Bromley
TITLE: 96-well Plate Growth Curve Setup AUTHORS: Norman van Rhijn, Panagiotis Papastamoulis, Takanori Furukawa, Magnus Rattray, Elaine Bignell, Mike Bromley [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Generally, growth assays for filamentous fungi have been performed on solid media, either...
["[Plate preparation]\nHarvest and dilute spores in PBS+0.01% Tween to 4*105 spores/mL.", "[Plate preparation]\nAliquot 5 uL of this spore stock per well of a CytoOne 96-well plate (Starlab) non-coated, this will be overlayed with 195 uL of liquid media to a total volume of 200 uL.\nThis equals to 2000 spores (absolute...