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94,844
Barcoded and targeted cDNA library preparation for Oxford Nanopore Technologies sequencing
4
dx.doi.org/10.17504/protocols.io.kqdg3xzwzg25/v1
https://www.protocols.io/view/barcoded-and-targeted-cdna-library-preparation-for-c8u4zwyw
Rosemary A Bamford, Szi Kay Leung, Aaron Jeffries, Jonathan Mill
TITLE: Barcoded and targeted cDNA library preparation for Oxford Nanopore Technologies sequencing AUTHORS: Rosemary A Bamford, Szi Kay Leung, Aaron Jeffries, Jonathan Mill [DESCRIPTION] The NEBNext Single Cell/Low Input cDNA Synthesis & Amplification Module (NEB) was adapted for the purpose of adding Oxford Nanopore ...
["[Bespoke barcoding of cDNA] Primer annealing for first strand synthesis\nPrepare in a PCR tube on ice. A reaction mix of up to 7 µL of total RNA (75 ng ) is added to 1 µL 2 millimolar (mM) TSO-ONT primer, 1 µL 10 millimolar (mM) dNTPs (NEB) and made up to 9 µL with nuclease-free H2O. Gently invert a few times ...
77,594
Single-cell dissociation of Drosophila melanogaster pupal tarsi
4
dx.doi.org/10.17504/protocols.io.x54v9dzbmg3e/v2
https://www.protocols.click/view/single-cell-dissociation-of-drosophila-melanogaste-cpz2vp8e
Ben R. Hopkins, Olga Barmina, Artyom Kopp
TITLE: Single-cell dissociation of Drosophila melanogaster pupal tarsi AUTHORS: Ben R. Hopkins, Olga Barmina, Artyom Kopp [DESCRIPTION] This protocol outlines a step-by-step guide to generating single-cell suspensions of Drosophila melanogaster pupal tarsi for use in 10x single-cell transcriptome profiling. This proto...
["[Collecting, sexing, and ageing pupae] Collect white prepupae. Individuals should meet the P1 aging criteria laid out by Bainbridge and Bownes (1981): the pupae should be white or cream coloured, have stopped moving completely, and display everted anterior spiracles.", "Fold a kimwipe in half and then in half again a...
null
null
null
dx.doi.org/10.17504/protocols.io.qxzdxp6
null
null
TITLE: No Title AUTHORS: [BEFORE_START] <p>Before starting, make 10X sterile Daigo solution in artificial seawater (ASW):</p> <p>Dissolve 2.56 g Daigo powder in 1L ASW and filter sterilize through a 0.22 µm filter.</p> [GUIDELINES] <p> </p> <p> </p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
53,511
Western blot protocol for detecting ATP10B in mouse/rat brain
1
dx.doi.org/10.17504/protocols.io.byhfpt3n
https://www.protocols.io/view/western-blot-protocol-for-detecting-atp10b-in-mous-byhfpt3n
María Sanchiz Calvo, eduard.bentea , Veerle Baekelandt
TITLE: Western blot protocol for detecting ATP10B in mouse/rat brain AUTHORS: María Sanchiz Calvo, eduard.bentea , Veerle Baekelandt [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for detection of ATP10B in rat and mouse brain tissue by Western blotting</div></div> [STEPS] ?. [Day 1] Prep...
["[Day 1]\nPrepare samples for Western blotting : proteins in of volume (adjust with milliQ water) of 6X SDS buffer + 10% of β-mercaptoethanol\n30 µg\n12 µl\n2.4 µl", "[Day 1]\nBoil the samples for at\n98 °C", "[Day 1]\nLoad samples, together with of mass marker (PageRuler™ Plus Prestained Protein Ladder) on a 3-8% ...
49,640
Measurement of dissolved black carbon in water via benzenepolycarboxylic acid (BPCA) oxidation and quantification using aqueous, inorganic, high-performance liquid chromatography
6
dx.doi.org/10.17504/protocols.io.buqgnvtw
https://www.protocols.io/view/measurement-of-dissolved-black-carbon-in-water-via-buqgnvtw
Riley Barton, Sasha Wagner
TITLE: Measurement of dissolved black carbon in water via benzenepolycarboxylic acid (BPCA) oxidation and quantification using aqueous, inorganic, high-performance liquid chromatography AUTHORS: Riley Barton, Sasha Wagner [DESCRIPTION] Dissolved black carbon (DBC) is the condensed aromatic portion of dissolved organ...
["[Solid Phase Extraction: Prepare and condition cartridges] The solid phase extraction (SPE) cartridges used in this protocol (Agilent Bond Elut PPL cartridge, 1g, 6 mL) is packed with a styrene-divinylbenzene polymer resin that recovers high proportions of dissolved organic matter (DOM) on a per-carbon basis. The res...
19,548
Enterovirus (EV) D68 real-time RT-PCR (EV-D68-TM2018)
null
dx.doi.org/10.17504/protocols.io.xb4fiqw
null
Ian Mackay, Judy Northill
TITLE: Enterovirus (EV) D68 real-time RT-PCR (EV-D68-TM2018) AUTHORS: Ian Mackay, Judy Northill [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol aims to amplify enterovirus (EV) D68 viruses but not other viruses.</div><div class = "text-block">This protocol is modified from a previously...
["[Oligonucleotide sequences]\nAB1NameSequence 5'-3'2EV-D68-For1TGTTYCCACGGTTGAAAAYAA3EV-D68-For2TTCCCACGGTTGAAARNYRAC4EV-D68-RevCAAGCTACACACGGGTTAGT5EV-D68-FAM-TM2018FAM - CCGTTAWCCGCTATAGTACTTCGAGAAACC - BHQ1MODIFICATIONS TO THE PUBLISHED ASSAY:Two modified forward primers, targeting the same region as the original a...
null
null
null
dx.doi.org/10.17504/protocols.io.etibeke
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <strong>Materials:</strong><br /><br />1.) host and viral DNAs<br />2.) 1X TE buffer (10 mM Tris, pH 8.0, 1 mM EDTA)<br />3.) 1 M NaOH<br />4.) 1 M Acetic acid<br />5.) nylon membrane, cut to 12.3 X16 cm (2 sheets)<br />6.) Whatman 3MM filter paper, cut 12.3X16 cm (2 sheets)<br /...
[]
31,983
Quantitative (q)PCR and Differential Expression Analysis
null
dx.doi.org/10.17504/protocols.io.bbgpijvn
null
Jeffrey Lewis, Amanda Scholes
TITLE: Quantitative (q)PCR and Differential Expression Analysis AUTHORS: Jeffrey Lewis, Amanda Scholes [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>A generalizable protocol for measuring relative changes in gene expression via qPCR (cDNA synthesis, qPCR primer optimization, and qPCR analysi...
["[qPCR Primer Design]\n1. Primer Design\nWe use Primer3 to design primers: http://biotools.umassmed.edu/bioapps/primer3_www.cgiPrimers are designed to have a Tm as close to 58°C as possible. This helps to ensure that primer annealing will be similar for all reactions. Primers should amplify a product of ~100 - 200 bp....
null
null
null
dx.doi.org/10.17504/protocols.io.rced2te
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Stains zebrafish larvae with Sudan Black B, which marks neutrophil granules.</p> [BEFORE_START] <p>You will need a rocker/nutator/rotator at 4ºC and at RT.</p> <p> </p> <p>Make both 1x and 10x stocks of PBS from powder packets, in water.</p> <p>Make 70% Ethanol solution.</p>...
[]
24,821
Diagnostic Restriction Digest
null
dx.doi.org/10.17504/protocols.io.4gvgtw6
null
Addgene The Nonprofit Plasmid Repository
TITLE: Diagnostic Restriction Digest AUTHORS: Addgene The Nonprofit Plasmid Repository [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol is for running a diagnostic restriction digest. To see the full abstract and other resources, visit the </span><a href="https://www.addgene.org/p...
["[Verifying Total Plasmid Size -OR- Insert and Backbone Size]\nThe simplest form of diagnostic digest is one in which you just want to verify that the plasmid that you have is the expected size, or that it is composed of a backbone and insert of expected sizes. This is frequently done after performing either PCR - or ...
39,906
SOLUTION -07 - RPMI/FBS 1%
3
dx.doi.org/10.17504/protocols.io.bi8akhse
https://www.protocols.io/view/solution-07-rpmi-fbs-1-bi8akhse
Marco Cosentino, Elisa Storelli, Alessandra Luini, Massimiliano Legnaro, Emanuela Rasini, Marco Ferrari, Franca Marino
TITLE: SOLUTION -07 - RPMI/FBS 1% AUTHORS: Marco Cosentino, Elisa Storelli, Alessandra Luini, Massimiliano Legnaro, Emanuela Rasini, Marco Ferrari, Franca Marino [STEPS]
[]
65,598
Natural killer cell depletion in vivo (mouse)
4
dx.doi.org/10.17504/protocols.io.rm7vzyezxlx1/v1
https://www.protocols.io/view/natural-killer-cell-depletion-in-vivo-mouse-cca6sshe
Goran Tomic
TITLE: Natural killer cell depletion in vivo (mouse) AUTHORS: Goran Tomic [DESCRIPTION] This protocol describes a validated procedure for antibody depletion of natural killer cells in mice (C57Bl/6J and 129SvEv strains), combined with the injection of cancer cells i.v. It is based on combining a few references and te...
["Mice will receive an i.p. injection of the antibody in 200 µl PBS (Day 1) e.g. Monday", "Mice will receive an i.p. injection of the antibody in 200 µl PBS (Day 4) e.g. Thursday", "Mice will receive tail vein injection of the cell suspension in 100 µl PBS (Day 5) e.g. Friday", "Mice will receive an i.p. injection of t...
55,577
Genotyping Arabidopsis T-DNA lines
1
dx.doi.org/10.17504/protocols.io.b2hzqb76
https://www.protocols.io/view/genotyping-arabidopsis-t-dna-lines-b2hzqb76
Lynn Doran, Steven J Burgess
TITLE: Genotyping Arabidopsis T-DNA lines AUTHORS: Lynn Doran, Steven J Burgess [DESCRIPTION] This protocol is used for genotyping Arabidopsis seedlings to test for the presence of a transfer DNA (T-DNA) insertion. By using two primer sets it is possible to determine whether a seedling is homozygous, heterozygous o...
["[Prepare primer working solution] Re-suspend lyophilized primers in dH2O to a stock concentration of 100 mM. Note: primer sequences can be obtained from SALK T-DNA express if you have the T-DNA accession number (http://signal.salk.edu/tdnaprimers.2.html)", "[Prepare primer working solution] Create a 10 mM working sol...
107,187
Populating metadata templates for NCBI submissions using PulseNet 2.0
1
dx.doi.org/10.17504/protocols.io.3byl4qn4ovo5/v2
https://www.protocols.io/view/populating-metadata-templates-for-ncbi-submissions-dkwt4xen
Ruth Timme, Maria Balkey, Tina Lusk Pfefer, Candace Hope Bias
TITLE: Populating metadata templates for NCBI submissions using PulseNet 2.0 AUTHORS: Ruth Timme, Maria Balkey, Tina Lusk Pfefer, Candace Hope Bias [DESCRIPTION] PURPOSE: to define the standard operating procedure for collecting isolate metadata using PulseNet 2.0 for submission of food/environmental isolates to NCBI...
["Metadata SampleSheet preparation \n\nPopulate the metadata spreadsheet form before uploading your sequencing run or linking NCBI sequencing records within PulseNet 2.0. \n\nPlease download the following metadata template, Guidance is included in the first tab. \n\n \n\nOnce you have filled out the template, import m...
null
null
null
dx.doi.org/10.17504/protocols.io.mjgc4jw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Dicentric Chromosomes Assay</p> [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
54,647
Protocol for ABC Immunohistochemistry and Quantifying Nerves
4
dx.doi.org/10.17504/protocols.io.eq2lypoymlx9/v1
https://www.protocols.io/view/protocol-for-abc-immunohistochemistry-and-quantify-bzkxp4xn
Donald Hoover
TITLE: Protocol for ABC Immunohistochemistry and Quantifying Nerves AUTHORS: Donald Hoover [DESCRIPTION] Cardiac tissue samples were obtained from human donors and sent to East Tennessee State University for processing. At ETSU, the tissue was dissected, sectioned with a cryostat, and stained for neuronal markers usin...
["[Immunostaining Day 1] Using the PAP Pen, carefully draw a water barrier circle around the tissue sections on the slide — allow this circle to dry for several seconds or up to approximately one minute.", "[Immunostaining Day 1] Rinse slides by placing them into a Coplin jar filled with PBS (pH 7.3 - 7.4): 4 x 5 min e...
null
null
null
dx.doi.org/10.17504/protocols.io.g7nbzme
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div class="page" title="Page 4"> <div class="layoutArea"> <div class="column"> <p>Infrared fluorescence detection with Odyssey Family Imaging Systems provides a quantitative two-color detection method for Western blots.</p> <p> </p> <p>This protocol is designed to help you achi...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.t4ceqsw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Sterilization and post-hatch care of gnotobiotic Periplaneta americana, as adapted from Doll et. al. (see below) Doll J, Trexler P, Reynolds L, Bernard G. 1963. The use of peracetic acid to obtain germfree invertebrate eggs for gnotobiotic studies. Am Midl Nat 69:231-239 [STEP...
["[Collect oothecae egg cases) from colony] Pulling egg cases directly off females' abdomens ensures the eggs have not been sitting in the litter a long time.\nThis also typically results in \"cleaner\" oothecae, with less excrement on the outside", "[Physically clean any visible/major excrement off of the outside of t...
71,097
MagAttract + Metapolyzyme metagenomic gDNA extraction from skin swabs
4
dx.doi.org/10.17504/protocols.io.q26g7yr19gwz/v2
https://www.protocols.io/view/magattract-metapolyzyme-metagenomic-gdna-extractio-chnzt5f6
Natalie Ring
TITLE: MagAttract + Metapolyzyme metagenomic gDNA extraction from skin swabs AUTHORS: Natalie Ring [DESCRIPTION] A protocol for the metagenomic extraction of bacterial DNA from skin swab samples (optimised using canine swabs), for use in a rapid diagnostics pipeline. At the end of the protocol, the DNA is cleaned up a...
["[Extended pre-lysis spin down] Bathe swab tip in 3 ml PBS in the swab tube for 10 minutes, with occasional vortexing. Remove swab from tube, squeezing the sides as you do.\n\n3 mL \n10 min", "[Extended pre-lysis spin down] Pellet 2x 1.5 ml aliquots of cell-PBS solution in 1.5 ml tubes by centrifuging at maximum speed...
40,076
In-silico analysis
3
dx.doi.org/10.17504/protocols.io.bjdkki4w
https://www.protocols.io/view/in-silico-analysis-bjdkki4w
avinash.kale
TITLE: In-silico analysis AUTHORS: avinash.kale [STEPS]
[]
32,307
Protocol Temephos Bioassay
1
dx.doi.org/10.17504/protocols.io.bbstinen
https://www.protocols.io/view/protocol-temephos-bioassay-bbstinen
Lara Ferrero Gomez
TITLE: Protocol Temephos Bioassay AUTHORS: Lara Ferrero Gomez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Performing bioassays to assess the larval susceptibility to Temephos</div></div> [STEPS]
[]
63,905
Oprah Winfrey ACV Gummies Shark Tank: Where To Buy It?
1
dx.doi.org/10.17504/protocols.io.8epv59my5g1b/v1
https://www.protocols.io/view/oprah-winfrey-acv-gummies-shark-tank-where-to-buy-cam9sc96
Shark Tank
TITLE: Oprah Winfrey ACV Gummies Shark Tank: Where To Buy It? AUTHORS: Shark Tank [DESCRIPTION] Oprah Winfrey ACV Gummies Shark Tank: Where To Buy It? [STEPS] SECTION: Oprah Winfrey ACV Gummies Shark Tank: Where To Buy It? Oprah Winfrey Keto – The Supplement for a Spare Shape and Slim Body! There&#x27;s a superb p...
["[Oprah Winfrey ACV Gummies Shark Tank: Where To Buy It? Oprah Winfrey Keto – The Supplement for a Spare Shape and Slim Body! There&#x27;s a superb product out then in the request that we all know by the name that goes as the Oprah Winfrey Keto. This blog is a complete in- depth analysis of this outstanding as well ...
24,048
Production of magnetic GFP affinity beads using a Camelid anti-GFP nanobody
null
dx.doi.org/10.17504/protocols.io.3qqgmvw
null
Nichanok Auevechanichkul, Ruth Lintermann, Lennart Wirthmueller
TITLE: Production of magnetic GFP affinity beads using a Camelid anti-GFP nanobody AUTHORS: Nichanok Auevechanichkul, Ruth Lintermann, Lennart Wirthmueller [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Our protocol for production of 'home-made' magnetic GFP affinity beads using a Camelid anti-GFP...
[]
19,677
Lysogeny Broth (1L)
null
dx.doi.org/10.17504/protocols.io.xf5fjq6
null
Brian Smith, baltrus@email.arizona.edu micro.tucson
TITLE: Lysogeny Broth (1L) AUTHORS: Brian Smith, baltrus@email.arizona.edu micro.tucson [STEPS] ?. [Fill container aproximately 750mL of H2O] Add stir bar and set to mix ?. [Add ingredients] [Bacto Tryptone] [Bacto Yeast Extract] [NaCl] ?. [Bring volume to 1L by adding H2O] Allow to mix again with stir bar. ?. [Autcla...
["[Fill container aproximately 750mL of H2O]\nAdd stir bar and set to mix", "[Add ingredients]\n[Bacto Tryptone]\n[Bacto Yeast Extract]\n[NaCl]", "[Bring volume to 1L by adding H2O]\nAllow to mix again with stir bar.", "[Autclave]\nAdd agar to container to be autclavedThen add media\n15 g", "[Autclave]\nAutclave at 121...
70,871
In vitro transcription of guide RNAs and 5&#39;-triphosphate removal
1
dx.doi.org/10.17504/protocols.io.n2bvjyp5vk5w/v14
https://www.protocols.io/view/in-vitro-transcription-of-guide-rnas-and-5-39-trip-chfxt3pn
Mark Dewitt, Julia Wong, Beeke Wienert, Moritz F Schlapansky, Eric Aird
TITLE: In vitro transcription of guide RNAs and 5&#39;-triphosphate removal AUTHORS: Mark Dewitt, Julia Wong, Beeke Wienert, Moritz F Schlapansky, Eric Aird [DESCRIPTION] sgRNA template assembly, in vitro T7 transcription, and sgRNA column cleanup to remove 5'-triphosphate groups [GUIDELINES] The primers used are: o...
["[Design sgRNA and order PCR oligos] Add the desired protospacer sequence to the T7FwdVarV2 oligo and order the oligo from your favorite oligonucleotide supplier. There are many programs available for protospacer design that attempt to optimize on- and/or off-target activity. Which program is most useful depends upon ...
95,481
Histological processing of octocoral tissue
4
dx.doi.org/10.17504/protocols.io.81wgbx38nlpk/v1
https://www.protocols.io/view/histological-processing-of-octocoral-tissue-c9gzz3x6
Maria Rakka, Gala G Edery, Marina Carreiro-Silva
TITLE: Histological processing of octocoral tissue AUTHORS: Maria Rakka, Gala G Edery, Marina Carreiro-Silva [DESCRIPTION] In recent years, there has been a significant focus on coral habitats, encompassing both shallow and deep ecosystems. Historically, most studies concentrated on scleractinian corals, however cont...
["[Fixation] Use 10% seawater formalin to fix the samples", "[Dissection] Using a scalpel and forceps, make an incision parallel to the coral axis. Carefully separate the tissue from the axis by using the forceps, and remove the axis completely.", "[Decalcification] Place tissue in eppendorfs (or other plastic/glass vi...
93,737
Gewebesammlung Frischgewebe Nephrektomie
1
null
https://www.protocols.io/view/gewebesammlung-frischgewebe-nephrektomie-c7shznb6
Annika Fendler, Bettina Ergün
TITLE: Gewebesammlung Frischgewebe Nephrektomie AUTHORS: Annika Fendler, Bettina Ergün [DESCRIPTION] Dieses Protokoll beschreibt die Schritte für die Sammlung von Frischgewebe, Gefriergewebe (Fresh-frozen), und Blut von Patienten mit Nierenzellkarzinom nach Nephrektomie. Verwandte Dokumente: Protokoll zur Blutaufa...
["[Gewebesammlung im Schnellschnitt] Nach Rückmeldung des Schnellschnitts geht eine Person zur Gewebeentnahme.  \nTüte Niere und Stickstoffbehälter mitnehmen.\nArbeitsanleitung (siehe Abbildung) beachten.", "[Gewebesammlung im Schnellschnitt]", "[Gewebesammlung im Schnellschnitt] 3x 15 ml Falcons mit 10,2 ml Transportm...
null
null
null
dx.doi.org/10.17504/protocols.io.fjwbkpe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>The printed instructions accompanying each Foldscope kit are very detailed and easy to follow. Even easier is <a href="https://www.youtube.com/watch?v=wJtJJixMncA" target="_blank">this</a> excellent 14-minute video tutorial for the assembly. </p> <p> </p> <p>I didn't know abo...
[]
66,943
DNA extraction (BOMB)
4
null
https://www.protocols.io/view/dna-extraction-bomb-cdk7s4zn
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 2ml enppendorf tube SECTION: Sample Collection 2. Add200 µL of 0.5mm beads to 2ml enppendorf tube SECTION: Sample Collection 3. Add 225 µL of TE...
["[Sample Collection] Add 200 µL of 1mm beads to 2ml enppendorf tube", "[Sample Collection] Add200 µL of 0.5mm beads to 2ml enppendorf tube", "[Sample Collection] Add 225 µL of TE buffer to 2ml enppendorf tube", "[Sample Collection] Add 375 µL of lysis buffer to 2ml enppendorf tube", "[Sample Collection] Add 267 µL of ...
79,924
Open, non-comparative, clinical investigation to evaluate the performance and safety of the medical device H42 (collagen paste for filling) in repairing periodontal pockets due to periodontitis.
1
dx.doi.org/10.17504/protocols.io.4r3l27rzxg1y/v2
https://www.protocols.io/view/open-non-comparative-clinical-investigation-to-eva-csauwaew
franco.barattini
TITLE: Open, non-comparative, clinical investigation to evaluate the performance and safety of the medical device H42 (collagen paste for filling) in repairing periodontal pockets due to periodontitis. AUTHORS: franco.barattini [DESCRIPTION] Research protocol to evaluate safety and overall performance of a collagen p...
["[Open, non-comparative, clinical investigation to evaluate the performance and safety of the medical device H42 (collagen paste for filling) in repairing periodontal pockets due to periodontitis.] Open, non-comparative, clinical investigation to evaluate the performance and safety of the medical device H42 (collagen ...
null
null
null
dx.doi.org/10.17504/protocols.io.dp95r5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Northeastern's method for plasmid extraction and purification. From Thermo Scientific's GeneJET Plasmid Miniprep Kit [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.cjrum5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This is the protocol for a large-scale Protein Expression Using BL21(DE3) Competent E. coli cells(C2527). [BEFORE_START] Determine the optimal time/temperature for the protein expression in a small scale trial. [GUIDELINES] <strong>BL21(DE3)&nbsp;Genotype:</strong><br />fhuA2 ...
[]
81,626
B-4 BLOOD TESTING
4
dx.doi.org/10.17504/protocols.io.rm7vzbze8vx1/v1
https://www.protocols.io/view/b-4-blood-testing-ctx2wpqe
REDI-NET Consortium
TITLE: B-4 BLOOD TESTING AUTHORS: REDI-NET Consortium [DESCRIPTION] This protocol details standard operating procedure for blood testing. [BEFORE_START] BEFORE START Check the DNA and RNA concentrations in each sample of total nucleic acid (TNA) extraction. If the concentrations are detectable, choose the sequencin...
["[gDNA PREPARATION] When the RNA concentration of the sample is lower than the detectable range of the Qubit High Sensitivity Assay (<0.01 ng/µl), the sample is subjected to gDNA sequencing. The cDNA synthesis can be skipped.", "[gDNA PREPARATION] When the DNA concentration >10 ng/µl, calculate the required volume of ...
null
null
null
dx.doi.org/10.17504/protocols.io.kvycw7w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Paraffin sections of formalin-fixed human pancreas and isolated human islets were treated with 0.05% pronase for antigen retrieval, blocked with 2% bovine serum albumin (BSA; Sigma)/phosphate buffered saline (PBS), incubated overnight (4°C) with 10E4 (anti-HS) mAb (1/10; US B...
[]
76,744
Setting up Zotero and Google Drive for syncing reference libraries (PC)
1
null
https://www.protocols.io/view/setting-up-zotero-and-google-drive-for-syncing-ref-cn7gvhjw
Carie M. Frantz
TITLE: Setting up Zotero and Google Drive for syncing reference libraries (PC) AUTHORS: Carie M. Frantz [DESCRIPTION] This protocol is useful if you wish to sync a LARGE reference library across multiple computers. It uses Zotero, which has built-in free syncing for up to 300 MB of files (roughly 50-100 papers). For l...
["[Set up Google Drive syncing] Install Drive for desktop on both computers", "[Install and set up Zotero on Computer 1] Install Zotero desktop\n\nDefault installation settings are fine.", "[Install and set up Zotero on Computer 2] Repeat the steps above for your second computer\n\nIf you do not have a prior (e.g., Men...
65,948
RNAlater recipe
4
dx.doi.org/10.17504/protocols.io.bp2l61w35vqe/v1
https://www.protocols.io/view/rnalater-recipe-ccm4su8w
Yin-Tse Huang
TITLE: RNAlater recipe AUTHORS: Yin-Tse Huang [DESCRIPTION] RNAlater recipe [STEPS] SECTION: Before you start 1. Submerge all your equipment and glassware in 1% bleach to get rid of nuclease SECTION: Reagents None. 0.5M Disodium dihydrate EDTA SECTION: Reagents None. SECTION: Before you start None. Autoclave ...
["[Before you start] Submerge all your equipment and glassware in 1% bleach to get rid of nuclease", "[Reagents] 0.5M Disodium dihydrate EDTA", "[Reagents]", "[Before you start] Autoclave to deactivate DEPC", "[Before you start] Keep the DEPC treated water at 4 °C", "[Reagents] DEPC treated water\n1. Add 1 mL DEPC in 1...
80,491
Forage and Range Research Laboratory Nursery and Plant Measurement Standard Operating Procedures (SOP) and Protocols
1
null
https://www.protocols.io/view/forage-and-range-research-laboratory-nursery-and-p-csujweun
blair.waldron, B Shaun Bushman, Alexander J Hernandez, Kevin B Jensen, Thomas A Jones, Steven R Larson, Thomas A Monaco, Michael D Peel, Matthew D Robbins, Joseph G. Robins, Richard R-C Wang
TITLE: Forage and Range Research Laboratory Nursery and Plant Measurement Standard Operating Procedures (SOP) and Protocols AUTHORS: blair.waldron, B Shaun Bushman, Alexander J Hernandez, Kevin B Jensen, Thomas A Jones, Steven R Larson, Thomas A Monaco, Michael D Peel, Matthew D Robbins, Joseph G. Robins, Richard R-C W...
["[Breeding Nursery and Plant Trait Measurements] Forage Mass", "[Breeding Nursery and Plant Trait Measurements] Using a sickle bar or flail forage harvester, cut forage plots to an 8-cm stubble height. On research sites unsuitable for the harvester, hand harvest the forage from at least three random 1-m2 quadrats thro...
45,040
Mollusk pedal mucus effects on epilithic biofilms
4
null
https://www.protocols.io/view/mollusk-pedal-mucus-effects-on-epilithic-biofilms-bp8qmrvw
Clara Maria Arboleda-Baena, Claudia Pareja , Isadora Pla, Ramiro Logares, Rodrigo De La Iglesia, Sergio A. Navarrete
TITLE: Mollusk pedal mucus effects on epilithic biofilms AUTHORS: Clara Maria Arboleda-Baena, Claudia Pareja , Isadora Pla, Ramiro Logares, Rodrigo De La Iglesia, Sergio A. Navarrete [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. We cultured epilithic biofilms on cover glass slides inside a Polycarbonate ...
["We cultured epilithic biofilms on cover glass slides inside a Polycarbonate cell culture plate of 6-Wells, for one week in K medium (Keller et al. 1987).", "We collected, during nocturnal low tides, animals of each mollusk species from wave-exposed platforms and brought them to the laboratory in coolers and then pla...
47,529
Bulk DNA Extraction (Ding Lab)
4
dx.doi.org/10.17504/protocols.io.bsnhndb6
https://www.protocols.io/view/bulk-dna-extraction-ding-lab-bsnhndb6
Reyka Jayasinghe, Li Ding, Feng Chen, Satok
TITLE: Bulk DNA Extraction (Ding Lab) AUTHORS: Reyka Jayasinghe, Li Ding, Feng Chen, Satok [DESCRIPTION] Bulk DNA Extraction [STEPS] 1. Cut tissue (~5 mg) and place in a 1.5 mL microcentrifuge tube. Add 180 µL Buffer ATL and 20 µL proteinase K, mix by vortexing and incubate at 56°C until completely lysed (1–3 h). 2. ...
["Cut tissue (~5 mg) and place in a 1.5 mL microcentrifuge tube. Add 180 µL Buffer ATL and 20 µL proteinase K, mix by vortexing and incubate at 56°C until completely lysed (1–3 h).", "Add 2 µL RNase (100 mg/ml) and incubate for 30 min. at 37°C", "Add 200 µL Buffer AL. Vortex for 15 sec.", "Incubate at 70°C for 10 min. ...
86,065
FM1-43 endocytic uptake assay in HIPSC derived neurons
4
dx.doi.org/10.17504/protocols.io.3byl4qj52vo5/v1
https://www.protocols.io/view/fm1-43-endocytic-uptake-assay-in-hipsc-derived-neu-cyarxsd6
Sakthi Kumar
TITLE: FM1-43 endocytic uptake assay in HIPSC derived neurons AUTHORS: Sakthi Kumar [DESCRIPTION] FM1-43 is a lipophilic styryl dye, nontoxic to the cells, virtually nonfluorescent in aqueous media. The dye will become fluorescent once it binds to the plasma membrane. In neurons that are actively releasing neurotransm...
["[Reagents needed:] FM1-43 dye (Thermo fisher scientific, catalog no: T3163).\nThis dye is available in both live and fixable versions. Also, available in different colors like FM4-64", "[Mature neurons- hipsc derived Dopaminergic, Cortical or GABAergic neurons:] mDA, CTX or GABAergic neuron progenitors were plated in...
null
null
null
dx.doi.org/10.17504/protocols.io.kwdcxa6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Intracavernous pressure (ICP) measurement is a well-established technique for assessing the erectile function, which was performed by cannulating either crus or shaft of the penis. But yet, as far as we know, no consensus has reached on the ES parameters of frequency and volt...
[]
94,545
Laptop setup and piranhaGUI install
5
null
https://www.protocols.io/view/laptop-setup-and-piranhagui-install-c8jrzum6
Aine.OToole
TITLE: Laptop setup and piranhaGUI install AUTHORS: Aine.OToole [DESCRIPTION] This protocol provides step-by-step guidance on installing piranhaGUI and its dependency Docker Desktop. [STEPS] SECTION: Install piranha through piranhaGUI 7. PiranhaGUI can be installed on a Windows, MacOSX or Linux machine. To install...
["[Install piranha through piranhaGUI] PiranhaGUI can be installed on a Windows, MacOSX or Linux machine. \nTo install piranhaGUI, navigate to piranhaGUI GitHub on a web browser. You should see a page similar to the one below, which shows the latest releases of piranha GUI, along with some installation instructions.", ...
78,418
Characterization of human immune cell subpopulations in cerebrospinal fluid using mass cytometry.
5
dx.doi.org/10.17504/protocols.io.36wgqjp4ovk5/v1
https://www.protocols.io/view/characterization-of-human-immune-cell-subpopulatio-cqtsvwne
Gerardina Gallaccio, Meng Wang, Stephan Schlickeiser, Desiree Kunkel, chotima.boettcher, Camila Fernández-Zapata
TITLE: Characterization of human immune cell subpopulations in cerebrospinal fluid using mass cytometry. AUTHORS: Gerardina Gallaccio, Meng Wang, Stephan Schlickeiser, Desiree Kunkel, chotima.boettcher, Camila Fernández-Zapata [DESCRIPTION] Phenotypic and compositional changes of immune cells in cerebrospinal fluid (...
["[Sample collection and storage] Prepare the anchor sample. \n\nAn anchor sample is peripheral blood mononuclear cells (PBMCs) used as internal reference across different measurements/batches to facilitate the signal normalization8.\nHowever, cell types other than PBMCs can also be used as an anchor sample but they sh...
50,879
OMS Atlas OCT Spatial Mapping
1
dx.doi.org/10.17504/protocols.io.bvw7n7hn
https://www.protocols.io/view/oms-atlas-oct-spatial-mapping-bvw7n7hn
Brett Johnson, Danielle Galipeau, Todd Camp, George Thomas
TITLE: OMS Atlas OCT Spatial Mapping AUTHORS: Brett Johnson, Danielle Galipeau, Todd Camp, George Thomas [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes the procedure by which the OMS Atlas serially sections an OCT block, prepares the resulting slides and samples, and then di...
["[Preparation]\nVerify the identity of the OCT block to be cut against written request for sectioning.", "[Preparation]\nRemove OCT block from freezer and acclimate to cryostat ( ) for minimum of .\n-80 °C\n-20 °C", "[Preparation]\nLabel all slides and cryotubes with a unique BEMS ID and Part#, corresponding to the w...
62,967
Botanical Farms CBD Gummies - Reviews, Side Effects, Benefits, Price &amp; Ingredients!
3
dx.doi.org/10.17504/protocols.io.4r3l2oyqpv1y/v1
https://www.protocols.io/view/botanical-farms-cbd-gummies-reviews-side-effects-b-b9qxr5xn
Botanical Farms CBD Gummies
TITLE: Botanical Farms CBD Gummies - Reviews, Side Effects, Benefits, Price &amp; Ingredients! AUTHORS: Botanical Farms CBD Gummies [DESCRIPTION] Botanical Farms CBD Gummies [STEPS]
[]
35,633
PhotoId-Whale: blue whale dorsal fin classification for mobile devices
1
dx.doi.org/10.17504/protocols.io.be2rjgd6
https://www.protocols.io/view/photoid-whale-blue-whale-dorsal-fin-classification-be2rjgd6
Rosa I Ramos-Arredondo, Blanca E, Diane Gendron, J. Francisco Gallegos-Funes, Dante Mújica-Vargas, J.B. Rosas-Fernández
TITLE: PhotoId-Whale: blue whale dorsal fin classification for mobile devices AUTHORS: Rosa I Ramos-Arredondo, Blanca E, Diane Gendron, J. Francisco Gallegos-Funes, Dante Mújica-Vargas, J.B. Rosas-Fernández [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Photoidentification is a method used by biolo...
["[Image to be classified]\nObtain the image to be classified. The image is obtained from the database or acquired in the field. CICIMAR-IPN researchers.", "[Image Pre-Processing]\nThis stage is divided into two substages, which are described belowi)\tROI selection: by means of a graphical interface, the initial point ...
49,171
MIBI staining
1
dx.doi.org/10.17504/protocols.io.bt9tnr6n
https://www.protocols.io/view/mibi-staining-bt9tnr6n
Marc Bosse, Sean Bendall, Mike Angelo
TITLE: MIBI staining AUTHORS: Marc Bosse, Sean Bendall, Mike Angelo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is the standard FFPE tissue staining procedure recommended for Multiplex Ion Beam Imaging Time of Flight instrument (MIBI_TOF) developed in the Sean C. Bendall and Michae...
["[Slide for MIBI]\nFFPE or frozen sections should be deposited on special conductive slides for MIBIIt is recommended to use freshly cut tissue sections. Otherwise tissue section slides should be stored properly using different state of the art methods (vacuum chamber, nitrogen chamber or vacuum sealed bags)", "[Slid...
25,813
SELECTION AND ENRICHMENT OF TRANSGENIC CELL POPULATIONS (Basic Protocol 3)
null
dx.doi.org/10.17504/protocols.io.5fvg3n6
null
Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward
TITLE: SELECTION AND ENRICHMENT OF TRANSGENIC CELL POPULATIONS (Basic Protocol 3) AUTHORS: Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>While lipid-mediated transient transfection and expressio...
["[Cell Preparation]\nCulture cells after transfection and expand to at least one full 6-well plate or one 10 cm dish by EDTA split (see Basic Protocol 1).\nPost-transfection recovery and expansion should take about 1 week; this enables the degradation / dilution of transient plasmids and permits expansion of edited ce...
null
null
null
dx.doi.org/10.17504/protocols.io.iykcfuw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.d7r9m5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <!-- [if gte mso 9]><xml> <o:OfficeDocumentSettings> <o:AllowPNG/> </o:OfficeDocumentSettings> </xml><![endif]--> <!-- [if gte mso 9]><xml> <w:WordDocument> <w:View>Normal</w:View> <w:Zoom>0</w:Zoom> <w:TrackMoves/> <w:TrackFormatting/> <w:PunctuationKerning/> <...
[]
46,867
Actinobacteria collection, enrichment and isolation
4
dx.doi.org/10.17504/protocols.io.brztm76n
https://www.protocols.io/view/actinobacteria-collection-enrichment-and-isolation-brztm76n
Rene Flores Clavo, Cristian Daniel Asmat Ortega, Nataly Ruiz Quinones
TITLE: Actinobacteria collection, enrichment and isolation AUTHORS: Rene Flores Clavo, Cristian Daniel Asmat Ortega, Nataly Ruiz Quinones [STEPS] ?. [Collection] Collect samples aseptically in 600 ml screw-capped glass jar. ?. [Collection] Pack the flasks in cool boxes with ice bags and transport them to the laborator...
["[Collection]\nCollect samples aseptically in 600 ml screw-capped glass jar.", "[Collection]\nPack the flasks in cool boxes with ice bags and transport them to the laboratory", "[Collection]\nIdentify each sample by writing a code, sampling date, depth, sample color, pH, and sample location", "[Enrichment and pre-trea...
100,443
Non-destructive microplastic isolation from water and sediment samples
0
dx.doi.org/10.17504/protocols.io.4r3l2q264l1y/v1
https://www.protocols.io/view/non-destructive-microplastic-isolation-from-water-deb33aqn
Jonas Stadfeld, Sneha Suresh, Srijak Bhatnagar
TITLE: Non-destructive microplastic isolation from water and sediment samples AUTHORS: Jonas Stadfeld, Sneha Suresh, Srijak Bhatnagar [DESCRIPTION] Microplastics in aquatic ecosystems serve as unique habitats for diverse microbial communities, collectively referred to as the plastisphere. Investigating these microbes ...
["[Microplastic Isolation - Water] NOTE: Use only metal and glass equipment. Tubing should be silicone.", "[Microplastic Isolation - Water] Filter at least 40L of water through a 20µm Cellulose filter (47 mm).\n\nNote: Depending upon the murkiness, the filter may clog before 40L is filtered. In that case, multiple filt...
48,735
Human Lung Digestion: Deriving a single cell suspension
4
null
https://www.protocols.io/view/human-lung-digestion-deriving-a-single-cell-suspen-btt7nnrn
Morrisey Lab
TITLE: Human Lung Digestion: Deriving a single cell suspension AUTHORS: Morrisey Lab [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Human Lung Digestion: Deriving a single cell suspension </span></div></div> [STEPS]
[]
48,141
Monitoring fly feeding behavior and timing by beetle luciferase reporters
1
dx.doi.org/10.17504/protocols.io.bs9mnh46
https://www.protocols.io/view/monitoring-fly-feeding-behavior-and-timing-by-beet-bs9mnh46
Misha Koksharov
TITLE: Monitoring fly feeding behavior and timing by beetle luciferase reporters AUTHORS: Misha Koksharov [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Feeding behavior of flies carrying a beetle (e.g. firefly) luciferase gene can be conveniently monitored in real-time by measuring bioluminescence...
["[Placing flies into 96-well plates]\nAnesthetize flies by cooling them on ice or by using a CO2 pad. The CO2 pads are the most common method used in fly labs while cooling is the simplest. \"In addition, it [cooling] is the only method which will not affect fly neurology, therefore behavior studies may begin after t...
28,660
RNA extraction with PGTX
null
dx.doi.org/10.17504/protocols.io.78uhrww
null
Lutz Berwanger
TITLE: RNA extraction with PGTX AUTHORS: Lutz Berwanger [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is used for RNA extraction in cyanobacteria after Pinto et al. 2009.</div><div class = "text-block"><span style = "font-weight:bold;">Pinto, Fernando Lopes; Thapper, Anders; Sontheim...
["Centrifuge for 3 minutes at 4˚C and 4.000 g.", "Resuspend the cell pellet in 1 ml of PGTX. Freeze in liquid nitrogen and store at -20 ˚C.\nWear goggles, a lab coat and gloves when dealing with PGTX and liquid nitrogen.", "Incubate for 5 min at 95 °C, shaking at 250 rpm in Thermomixer (Eppendorf)", "Rapidly chill 5 mi...
null
null
null
dx.doi.org/10.17504/protocols.io.k7yczpw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Proximity Ligation Protocol with Concurrent IF</p> [STEPS]
[]
58,739
96 well spin column TNA extraction from plants - CTAB method
4
dx.doi.org/10.17504/protocols.io.n2bvj685plk5/v1
https://www.protocols.io/view/96-well-spin-column-tna-extraction-from-plants-cta-b5ktq4wn
James JN JN Kitson
TITLE: 96 well spin column TNA extraction from plants - CTAB method AUTHORS: James JN JN Kitson [DESCRIPTION] This protocol is designed for extracting total nucleic acids (TNA) from plant material. In reality the drying step probably means that you won't isolate plant mRNA but viral RNA (and probably plant ribosomal R...
["[Preparation of grinding tubes and buffer] Source steel beads (ball bearings) for tissue grinding (Tungsten beads are not usually necessary). We use hardened 3mm carbon steel or stainless steel bearings from simplybearings.co.uk. This protocol requires three beads per sample tube.", "[Initial homogenisation of plant ...
57,047
Intracellular cytokine detection based on flow cytometry in hemocytes from Galleria mellonella larvae
4
dx.doi.org/10.17504/protocols.io.b3xxqppn
https://www.protocols.io/view/intracellular-cytokine-detection-based-on-flow-cyt-b3xxqppn
Anna Katarzyna Wrońska, Agata Kaczmarek, Mieczysława Irena Boguś
TITLE: Intracellular cytokine detection based on flow cytometry in hemocytes from Galleria mellonella larvae AUTHORS: Anna Katarzyna Wrońska, Agata Kaczmarek, Mieczysława Irena Boguś [DESCRIPTION] Invertebrates are becoming increasingly popular models for research on the immune system. The innate immunity possessed by...
["[Sample preparation] Before bleeding, wash the larvae with distilled water (15 seconds) and then immerse briefly (5 seconds) in 70% (v/v) ethanol to sterilize their surfaces, thus reducing the chance of contamination of hemolymph samples.", "[Sample preparation] Collect the hemolymph into sterile tube from the larvae...
null
null
null
dx.doi.org/10.17504/protocols.io.mepc3dn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluoresc...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.tfeejje
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ssdeea6
null
null
TITLE: No Title AUTHORS: [BEFORE_START] <p>Clean the work area with 70% alcohol. Use all filters and autoclaved tips. Refrigerate your centrifuge to 4C.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ib6care
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Please contact Dr. Steven Wilhelm (wilhelm@utk.edu) for additional information regarding this protocol.</p> <p> </p> <p>Modified from <span style="font-family: 'Segoe UI', sans-serif; font-size: small;">Schwyn, B. &amp; Neilands, J. B. Universal chemical assay for the detecti...
[]
71,297
Transfection of Atlantic salmon primary hepatocytes
4
dx.doi.org/10.17504/protocols.io.j8nlkw4p1l5r/v1
https://www.protocols.io/view/transfection-of-atlantic-salmon-primary-hepatocyte-chu9t6z6
Alex K. Datsomor, Ragnhild Wilberg, Jacob S. Torgersen, Simen R. Sandve, Thomas N Harvey
TITLE: Transfection of Atlantic salmon primary hepatocytes AUTHORS: Alex K. Datsomor, Ragnhild Wilberg, Jacob S. Torgersen, Simen R. Sandve, Thomas N Harvey [DESCRIPTION] This protocol is for isolation and transfection of primary hepatocytes from Atlantic salmon. We have confirmed plasmid transfection efficiency of up...
["[Prepare buffers and plates] Coat wells of the culture plate with 1x PEI.", "[Prepare buffers and plates] Dilute PEI stock 1:100 in 0.1 M borate buffer.", "[Prepare buffers and plates] Add sufficient volume to empty wells. Enough to cover the bottom of the well.", "[Prepare buffers and plates] Incubate for 60 min at ...
null
null
null
dx.doi.org/10.17504/protocols.io.et2beqe
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
82,751
immunofluorescent staining with anti-GFP and anti-CD63 antibodies
4
dx.doi.org/10.17504/protocols.io.14egn2pypg5d/v1
https://www.protocols.click/view/immunofluorescent-staining-with-anti-gfp-and-anti-cu27wyhn
rosanne.wouters, Peter Vangheluwe
TITLE: immunofluorescent staining with anti-GFP and anti-CD63 antibodies AUTHORS: rosanne.wouters, Peter Vangheluwe [DESCRIPTION] This protocol was used for immunofluorescent staining in fixed HeLa cells with anti-GFP and anti-CD63 antibodies, followed by confocal imaging. [STEPS] 2. fix cells with 4% paraformaldehyd...
["fix cells with 4% paraformaldehyde for 20 min at room temperature", "permeabilize cells with 0.1% Triton X-100 in PBS for 5 min", "block for 1 h with blocking buffer (PBS with 0.5% Tween20, 0.1% BSA, 0.2% FBS)", "incubate coverslips with primary antibodies for 2 h at room temperature\n(anti-CD63, exbio, 11-343-C100, ...
null
null
null
dx.doi.org/10.17504/protocols.io.c7bzim
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] . [STEPS] ?.
[]
88,645
Barcode Composition by Overlap-Extension PCR
1
null
https://www.protocols.io/view/barcode-composition-by-overlap-extension-pcr-c2tdyei6
Mathew Chu
TITLE: Barcode Composition by Overlap-Extension PCR AUTHORS: Mathew Chu [DESCRIPTION] Traditionally, DNA barcodes are synthesised as random oligonucleotides. However, this leads to uncertainty regarding the ground truth of barcode sequences in the experimental setting. Without reference sequences, it is impossible to ...
["[Single Stranded DNA Pools for Combinatorial Assembly] ssDNA oligos for combinatorial assembly can be ordered as a pool (oPool). For a final barcode of n units, each with m diversity, order a set of m different barcode sequences for each unit:\n \nwhere unit i (1 ≤ i ≤ n) consists of m barcodes flanked by left (L) an...
97,912
The State of Research on LGBTQ+ Fertility: Trends, Gaps, and Future Directions
0
dx.doi.org/10.17504/protocols.io.yxmvme7dbg3p/v1
https://www.protocols.io/view/the-state-of-research-on-lgbtq-fertility-trends-ga-dbuy2nxw
Demian Glujovsky, Julieta quaglia, Fiamma Belén Di Biase, Mariana Miguens, Romina Pesce, Belen Herrero, Agustín Ciapponi
TITLE: The State of Research on LGBTQ+ Fertility: Trends, Gaps, and Future Directions AUTHORS: Demian Glujovsky, Julieta quaglia, Fiamma Belén Di Biase, Mariana Miguens, Romina Pesce, Belen Herrero, Agustín Ciapponi [DESCRIPTION] The hypothesis is that in the last decade the trend of published studies about reproducti...
["[Background] Rationale and potential applied impact \nThe LGBTQ+ community has made significant progress in recent years in terms of visibility and awareness. This has probably contributed to an increase in the number of LGBTQ+ people who are seeking fertility treatments, gaining access to these types of treatments t...
null
null
null
dx.doi.org/10.17504/protocols.io.ukgeutw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] The following protocol describes resuspension of lyophilized primers from (4) 96-Well plates shipped from Integrated DNA Technologies (IDT) at 3nmol per well using the epMotion 5075. For information about ordering consult the Earth Micriobiome Projects website. Barcoded primer...
["[Prepare reagents] {\"blocks\":[{\"key\":\"9h1q2\",\"text\":\"In a sterile 30mL reservoir, add 11.725\\u00a0ml (required minimum volume) of PCR Clean Water to resuspend (4) 96-Well plates. \",\"type\":\"unstyled\",\"depth\":0,\"inlineStyleRanges\":[],\"entityRanges\":[],\"data\":[]},{\"key\":\"4odgr\",\"text\":\" \",...
72,557
NARMS-KS-1-Autoclave Use
4
null
https://www.protocols.io/view/narms-ks-1-autoclave-use-ci4mugu6
tdoerks
TITLE: NARMS-KS-1-Autoclave Use AUTHORS: tdoerks [DESCRIPTION] Instructions on how to use autoclave in Mosier P209. [STEPS] SECTION: Waste 1. Make sure waste to be autoclaved is in a biohazard bag (no more than 3/4 full) and place in autoclavable container. SECTION: Waste 2. Wrap 2 Steam Migrating Integrator Strips t...
["[Waste] Make sure waste to be autoclaved is in a biohazard bag (no more than 3/4 full) and place in autoclavable container.", "[Waste] Wrap 2 Steam Migrating Integrator Strips together with autoclave tape and a piece of string and place in the middle of the bag of waste to be autoclaved with the piece of string hangi...
40,094
Lyophilized metal-antibody reconstitution
4
dx.doi.org/10.17504/protocols.io.bjd6ki9e
https://www.protocols.io/view/lyophilized-metal-antibody-reconstitution-bjd6ki9e
Marc Bosse, Mike Angelo, Sean Bendall
TITLE: Lyophilized metal-antibody reconstitution AUTHORS: Marc Bosse, Mike Angelo, Sean Bendall [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes the procedure to prepare the reconstitution buffer and the resuspension of metal-labeled antibody.</div></div> [STEPS] ?. [Stock so...
["[Stock solutions preparation]\nPrepare Tris (10 mM), sodium azide (0.02%)", "[Stock solutions preparation]\nAdd Tris 1M, pH 7.5 in tube\n500 µl\n50 mL", "[Stock solutions preparation]\nAdd sodium azide ()\n500 µl", "[Stock solutions preparation]\nComplete with 50 mL ultrapure water", "[Stock solutions preparation]\...
97,346
Protein Digestion with S-trap Spin Columns using Conditioned Concentrated Media
4
dx.doi.org/10.17504/protocols.io.x54v928eml3e/v1
https://www.protocols.io/view/protein-digestion-with-s-trap-spin-columns-using-c-dbba2iie
Joanna Bons, J P Rose, M A Watson, B Schilling
TITLE: Protein Digestion with S-trap Spin Columns using Conditioned Concentrated Media AUTHORS: Joanna Bons, J P Rose, M A Watson, B Schilling [DESCRIPTION] Trypsin digestion of isolated proteins using S-trap Spin columns in preparation for downstream proteomic profiling. For trypsin digestion of proteins from condi...
["In a 2-mL microcentrifuge tube add an appropriate proportion of volume from the concentrated condition media, 10% SDS for a final concentration of 4% SDS, 1M TEAB pH 8 solution for a final concentration of 50 mM TEAB, and HPLC-grade water if necessary to bring the volume up to a minimum of 50 µL.", "Add 250 mM DTT fo...
57,083
Test for blog post
1
dx.doi.org/10.17504/protocols.io.n92ldz9y7v5b/v1
https://www.protocols.io/view/test-for-blog-post-b3y3qpyn
Emma Ganley
TITLE: Test for blog post AUTHORS: Emma Ganley [DESCRIPTION] Just a test [STEPS] 1. 2.
[]
60,115
Wastewater QC workflow in GalaxyTrakr (SSQuAWK3)
1
dx.doi.org/10.17504/protocols.io.kxygxzk5dv8j/v4
https://www.protocols.io/view/wastewater-qc-workflow-in-galaxytrakr-ssquawk3-b6xtrfnn
Jasmine Amirzadegan, Tunc Kayikcioglu, hugh.rand , Ruth Timme, Maria Balkey
TITLE: Wastewater QC workflow in GalaxyTrakr (SSQuAWK3) AUTHORS: Jasmine Amirzadegan, Tunc Kayikcioglu, hugh.rand , Ruth Timme, Maria Balkey [DESCRIPTION] PURPOSE: Step-by-step instructions for checking sequence quality for SARS-CoV-2 wastewater samples using SSQuAWK3: SARS - CoV - 2 Sequence Quality Assuranc...
["[Account set up] Create a GalaxyTrakr account here: https://account.galaxytrakr.org/Account/Register", "[Account set up] Log into your GalaxyTrakr account: https://galaxytrakr.org", "[Create a new history] Create a new history. \n\nWe recommend creating a new history for each new MiSeq sequence set with details and d...
21,332
Yale - HDL Cholesterol
null
dx.doi.org/10.17504/protocols.io.y3ufynw
null
John Stack, Gary Cline
TITLE: Yale - HDL Cholesterol AUTHORS: John Stack, Gary Cline [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">Procedure used to determine the concentration of HDL cholesterol in blood, serum, and plasma. HDL Cholestero...
["Calibrate Cobas for HDL analysis by running a lipid calibrator, HDL Direct Reagent Reagent 1 and HDL Direct Reagent 2.", "Sample handling as performed by the Cobas Mira Plus. a) Pipette 3µL of sample into cuvette. b) Add 180 µL of Direct Reagent 1. c) Add 60 µL of Direct Reagent 2. d) Mixture is incubated at 37ºC...
39,615
Open Source Microfluidic Scaffolds
5
dx.doi.org/10.17504/protocols.io.biw7kfhn
https://www.protocols.io/view/open-source-microfluidic-scaffolds-biw7kfhn
Harry Felton, Robert Hughes
TITLE: Open Source Microfluidic Scaffolds AUTHORS: Harry Felton, Robert Hughes [STEPS] ?. [Design] User designs the required microfluid system from the sub-systems available. ?. [Generate Models] Download, install and open Fusion 360. NB: Although Fusion 360 works on iOS and Android devices the add-in will not and so ...
["[Design]\nUser designs the required microfluid system from the sub-systems available.", "[Generate Models]\nDownload, install and open Fusion 360.\nNB: Although Fusion 360 works on iOS and Android devices the add-in will not and so a Windows or MacOS device should be used.", "[Generate Models]\nLoad the µ-fluid add-i...
77,719
In situ IRF4 detection in formalin-fixed, paraffin-embedded (FFPE) pig tissues
4
dx.doi.org/10.17504/protocols.io.4r3l27bdqg1y/v1
https://www.protocols.io/view/in-situ-irf4-detection-in-formalin-fixed-paraffin-cp5xvq7n
Jayne E Wiarda, Crystal Loving
TITLE: In situ IRF4 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 IRF4 [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 ...
44,817
Lab 6 Notebook
3
dx.doi.org/10.17504/protocols.io.bpzrmp56
https://www.protocols.io/view/lab-6-notebook-bpzrmp56
TITLE: Lab 6 Notebook AUTHORS: [STEPS]
[]
89,665
Pole Test
1
dx.doi.org/10.17504/protocols.io.3byl4q362vo5/v1
https://www.protocols.io/view/pole-test-c3s9ynh6
Jhodi Webster
TITLE: Pole Test AUTHORS: Jhodi Webster [DESCRIPTION] This protocol describes the pole test, a simple and rapid behavioral assay used to evaluate motor coordination, balance, and bradykinesia (slowness of movement) in mice and rats. The test involves placing an animal head-up at the top of a vertical textured pole and...
["[Setup] Allow for one hour habituation period for mice in the testing room.", "[Setup] Place pole (approx 20.5in long, with rubber bands place at 1 inch intervals)", "[Setup] Have an extra empty cage handy", "[Training:] place mouse nose-down at the base of the pole and allow it to climb off onto the floor of the enc...
101,203
Cell counting
0
dx.doi.org/10.17504/protocols.io.14egn66rql5d/v1
https://www.protocols.io/view/cell-counting-de3t3gnn
daniel.dautan daniel, Per Svenningsson
TITLE: Cell counting AUTHORS: daniel.dautan daniel, Per Svenningsson [DESCRIPTION] Used for counting cells labeled with immunofluorescent markers in mouse brain sections. Sections should be stained, mounted, and imaged with high resolution (2048 x 2048 scanning). [STEPS] 1. Using 2-3 sections to encompass the brain r...
["Using 2-3 sections to encompass the brain region of interest, acquire high resolution images (2048 x 2048) using tile scanning and z-stack acquisition.", "Import images into ImageJ.", "Apply maximum average projection and color adjust as needed, making sure to apply the same settings to all images.", "For each image,...
60,038
Screening whole proteome of Aedes aegypti and identification of potential targets for in-silico molecular and structural interaction studies against natural bioactives
5
dx.doi.org/10.17504/protocols.io.e6nvwkbj2vmk/v1
https://www.protocols.io/view/screening-whole-proteome-of-aedes-aegypti-and-iden-b6vere3e
Anagha S Setlur, Chandrashekar K, Manas Sarkar, Vidya Niranjan
TITLE: Screening whole proteome of Aedes aegypti and identification of potential targets for in-silico molecular and structural interaction studies against natural bioactives AUTHORS: Anagha S Setlur, Chandrashekar K, Manas Sarkar, Vidya Niranjan [DESCRIPTION] As a dangerous etiological agent for dengue, chik...
["[IDENTIFICATION OF PROTEIN TARGETS] Reference proteome identification\n\nAedes aegypti is a yellow fever mosquito, that is also an etiological agent for dengue, chikungunya and zika. Hence, identification of the significant proteins in the organism’s reference proteome by a whole proteome screening is essential to us...
72,274
Selecting a Region of Interest for Hidden Markov Modeling using ebFRET
1
null
https://www.protocols.io/view/selecting-a-region-of-interest-for-hidden-markov-m-citsuene
Clark Fritsch
TITLE: Selecting a Region of Interest for Hidden Markov Modeling using ebFRET AUTHORS: Clark Fritsch [DESCRIPTION] This protocol follows from the "Basic Analysis Protocol" and is the first step towards analyzing your data using Hidden Markov Modeling. In this protocol, you select traces containing features that you wi...
["After generating a catalog of traces that you consider worthy of further analysis and saving these traces to your \"GoodOnes.txt\" file for a given directory (as described in the \"Basic Analysis Protocol\") you will have the opportunity to measure specific parameters in your traces by manually selecting regions of i...
67,378
Ready XL Male Enhancement Reviews Pills Side Effects
1
dx.doi.org/10.17504/protocols.io.261genmqwg47/v1
https://www.protocols.io/view/ready-xl-male-enhancement-reviews-pills-side-effec-cd2ss8ee
Alpha State Male Enhancement
TITLE: Ready XL Male Enhancement Reviews Pills Side Effects AUTHORS: Alpha State Male Enhancement [DESCRIPTION] Ready XL Male Enhancement Reviews Pills Side Effects [STEPS] SECTION: Ready XL Male Enhancement Reviews Pills Side Effects Ready Xl Male Enhancement- Women blame individualities for being occupied with co...
["[Ready XL Male Enhancement Reviews Pills Side Effects Ready Xl Male Enhancement- Women blame individualities for being occupied with coitus. What a nationalist and disparaging comment! In any case, they are correct, so we should assume the stylish about them. Assuming that your abettor is womanish, she presumably ant...
95,635
Descending Platform
4
dx.doi.org/10.17504/protocols.io.14egn3ko6l5d/v2
https://www.protocols.io/view/descending-platform-c9mtz46n
daniel.dautan daniel, Per Svenningsson
TITLE: Descending Platform AUTHORS: daniel.dautan daniel, Per Svenningsson [DESCRIPTION] Behavioral test to assess motor function. The test is using a custom made 45-degree grid path with a width of 5cm and a length of 45cm. The 15cm one side is oriented with a 45-degree angle. The grid is formed by metal mesh with a ...
["Place the custom-made, metal mesh grid to have an angle of 45 degrees with the floor.", "At the end of the platform, place the animal cage to provide a safe target.", "Place a camera above the grid to record the entire procedure. Here we use the Logitech C920 webcam and windows camera software.", "Using the video, re...
30,602
Micro-CT imaging of iodine-stained rat stomach
1
dx.doi.org/10.17504/protocols.io.95ih84e
https://www.protocols.io/view/micro-ct-imaging-of-iodine-stained-rat-stomach-95ih84e
Deborah Jaffey, Terry Powley, Logan Chesney
TITLE: Micro-CT imaging of iodine-stained rat stomach AUTHORS: Deborah Jaffey, Terry Powley, Logan Chesney [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol describes how to prepare a rat stomach for micro-CT imaging to enable a 3D description of the organ. Iodine is used to stain the ti...
["[Training]\nSelect 4 rats within the preferred weight distribution. Each animal is trained to consume a quantity of palatable In the first 2 days, the animal is supplied with both regular rat chow and a half container of Dietgel (the Dietgel is put in a dish in the cage at 11AM) to accustom itself to the Dietgel. In...
36,525
Short protocol for mitochondrial CO1 gene analysis of shark
1
dx.doi.org/10.17504/protocols.io.bfwmjpc6
https://www.protocols.io/view/short-protocol-for-mitochondrial-co1-gene-analysis-bfwmjpc6
Sutanto Hadi, Noviar Andayani, Effin Muttaqin, Benaya M Simeon, Muhammad Ichsan, Beginer Subhan, Hawis Madduppa
TITLE: Short protocol for mitochondrial CO1 gene analysis of shark AUTHORS: Sutanto Hadi, Noviar Andayani, Effin Muttaqin, Benaya M Simeon, Muhammad Ichsan, Beginer Subhan, Hawis Madduppa [STEPS] ?. [Shark Tissue Sampling] Shark Tissue Sampling .justify:after { content: ""; display:inlin...
["[Shark Tissue Sampling]\nShark Tissue Sampling\n\t\t\t\t\t\t\t .justify:after {\n\t\t\t\t\t\t\t content: \"\";\n\t\t\t\t\t\t\t display:inline-block;\n\t\t\t\t\t\t\t width: 100%;\n\t\t\t\t\t\t\t }\n\t\t\t\t\t\t\tPrepare sampling tools and equipment: camera, ruler or scale band, gloves, scissors or cutter an...
102,397
Lysosomal activity DQ-Red BSA assay
0
dx.doi.org/10.17504/protocols.io.j8nlk8dp6l5r/v1
https://www.protocols.io/view/lysosomal-activity-dq-red-bsa-assay-df853ry6
Razaul Karim
TITLE: Lysosomal activity DQ-Red BSA assay AUTHORS: Razaul Karim [DESCRIPTION] This protocol details the Lysosomal activity DQ-Red BSA assay. [STEPS] SECTION: Cell Culture/Treatments: 1. 0.5x10^4/ml of cells plated on coverslips. SECTION: Cell Culture/Treatments: 2. PFF-488 treatment. SECTION: Cell Culture/Treatments...
["[Cell Culture/Treatments:] 0.5x10^4/ml of cells plated on coverslips.", "[Cell Culture/Treatments:] PFF-488 treatment.", "[Cell Culture/Treatments:] PBS/trypsin (0.01%) wash.", "[Cell Culture/Treatments:] DQ-BSA Red treatment: \n\n10 152790 min at 37 °C.", "[Cell Culture/Treatments:] Fix cells using 4% PFA.", "[Imagi...
92,203
Establishment of patient-derived enteroids/colonoids from endoscopic biopsies
4
dx.doi.org/10.17504/protocols.io.14egn3rxql5d/v1
https://www.protocols.io/view/establishment-of-patient-derived-enteroids-colonoi-c6ajzacn
Tatiana Karakasheva
TITLE: Establishment of patient-derived enteroids/colonoids from endoscopic biopsies AUTHORS: Tatiana Karakasheva [DESCRIPTION] This protocol describes the full procedure of generating an enteroid or colonoid culture from endoscopic biopsy (either freshly procured or recovered from cryopreservation), was developed for...
[]
71,186
Microscopy-based pUb-coverage measurements of mitochondria in iNeurons
4
dx.doi.org/10.17504/protocols.io.e6nvwj529lmk/v1
https://www.protocols.io/view/microscopy-based-pub-coverage-measurements-of-mito-chrst56e
Felix Kraus
TITLE: Microscopy-based pUb-coverage measurements of mitochondria in iNeurons AUTHORS: Felix Kraus [DESCRIPTION] Protocol for microscopy-based pUb-coverage measurements of mitochondria in iNeurons [STEPS] SECTION: Differentiation of iNeurons 22. Day 0: Treat AAVS1-TRE3G-NGN2 cells with Accutase and plate the dissocia...
["[Differentiation of iNeurons] Day 0: Treat AAVS1-TRE3G-NGN2 cells with Accutase and plate the dissociated cells in matrigel-coated 6-well plates (2x105 cells/well) in ND1 Medium supplemented with Y27632 (10 μM). \nND1 Medium: \nDMEM/F12 \nN2 (100x) 1x \nBDNF 10 ng/ml \nNT3 10 n...
98,503
SenNet URMC 10X Genomics Single-Nucleus RNA-Sequencing for Transcriptomic Profiling
1
dx.doi.org/10.17504/protocols.io.14egn6zb6l5d/v1
https://www.protocols.io/view/sennet-urmc-10x-genomics-single-nucleus-rna-sequen-dcff2tjn
Jeffrey Malik, blake, Gloria S Pryhuber
TITLE: SenNet URMC 10X Genomics Single-Nucleus RNA-Sequencing for Transcriptomic Profiling AUTHORS: Jeffrey Malik, blake, Gloria S Pryhuber [DESCRIPTION] 10X Genomics Single Cell 3' (v3.1) RNA sequencing is a microdroplet-based method that permits the effective capture and sequencing of the mRNA and pre-mRNA molecule...
["[Isolate Nuclei] Resuspend nuclei in 100 µL to 1 mL of PBS + 0.1% RNase Inhibitor (volume depends on target concentration)", "[Isolate Nuclei] Perform manual trypan blue nuclear exclusion counts on a hemacytometer.", "[Isolate Nuclei] Check nuclei integrity by light microscope concurrent with manual trypan exclusion ...
37,914
UABMC - Stemness Determination Protocol
4
null
https://www.protocols.io/view/uabmc-stemness-determination-protocol-bg92jz8e
Christopher Willey
TITLE: UABMC - Stemness Determination Protocol AUTHORS: Christopher Willey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This procedure describes the assays for determining stem-ness in the PDMC models. </div><div class = "text-block">The phenotypic and marker-based assays that can and will be use...
["[LIMITING DILUTION ASSAY]\nEnsure cultures are healthy with minimal percentages of dead cells", "[LIMITING DILUTION ASSAY]\nDissociate cells using Accutase as usual", "[LIMITING DILUTION ASSAY]\nWhile cells are dissociating, add stem cell media to inner wells (6x10) at 200uL per well and to well H2. Add water to the...
55,703
Isolation and RNA extraction from Marine Protist Single Cells for Transcriptomics (sc-RNAseq)
1
dx.doi.org/10.17504/protocols.io.x54v9mk3qg3e/v4
https://www.protocols.io/view/isolation-and-rna-extraction-from-marine-protist-s-b2mxqc7n
Sarah Romac
TITLE: Isolation and RNA extraction from Marine Protist Single Cells for Transcriptomics (sc-RNAseq) AUTHORS: Sarah Romac [DESCRIPTION] It is difficult to get clean RNA from uncultivated protists (phyto and zooplankton), as they have to be isolated by micromanipulation and very sensitive to exogenous contaminations. H...
["[1. Cell Isolation] Isolate individually protist cells (5-500µm in length) using a glass bent micropipette (or a micropipette adapted to the length of the microorganism) under a binocular microscope or an inverted microscope.", "[1. Cell Isolation] Wash each cell in three successive baths of 0.22µm-filtered and...
53,259
Female reproductive organs procurement
4
dx.doi.org/10.17504/protocols.io.bx9jpr4n
https://www.protocols.io/view/female-reproductive-organs-procurement-bx9jpr4n
kone
TITLE: Female reproductive organs procurement AUTHORS: kone [DESCRIPTION] Description of procurement of non-pregnant female reproductive tract from deceased donor for HuBMAP. [STEPS] SECTION: Modification of donor procurement 1. During the typical aortic cannulation in a standard abdominal organ procurement, the in...
["[Modification of donor procurement] During the typical aortic cannulation in a standard abdominal organ procurement, the inferior aorta distal to the cannulation site is ligated. This occurs usually at the level of the bifurcation of the aorta with the common iliac arteries. This ligation is necessary in order to p...
null
null
null
dx.doi.org/10.17504/protocols.io.nyzdfx6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This is the protocol for the touchdown PCR assay</p> [BEFORE_START] <p>The assay has been optimized and validated for the Bio-rad C1000 Touch Thermocycler.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
103,409
comments retest discarded 
0
null
https://www.protocols.io/view/comments-retest-discarded-dg8r3zv6
Maria Gulyakina
TITLE: comments retest discarded  AUTHORS: Maria Gulyakina [DESCRIPTION] qa [STEPS] 1. test 2. comment 3. test 2 4. test 3
["test", "comment", "test 2", "test 3"]
66,639
Extracellular DNA extraction from lake sediments
1
null
https://www.protocols.io/view/extracellular-dna-extraction-from-lake-sediments-cdbps2mn
Charline Giguet-Covex, Pierre Taberlet, Francesco Gentile Ficetola
TITLE: Extracellular DNA extraction from lake sediments AUTHORS: Charline Giguet-Covex, Pierre Taberlet, Francesco Gentile Ficetola [DESCRIPTION] Over the past decade, an increasing number of studies has used environmental DNA from lake sediments to trace past lake ecosystem and landscape changes, agricultural activ...
["[exDNA extraction] Preparation of the SW2 buffer within the kit (NucleoSpin® Soil kit from Macherey-Nagel):\n\nThis step is only FOR the FIRST USE of this BUFFER\n\nAdd correct volume of ethanol to SW2 (e.g. for 250 prep kit, add 400 mLof ethanol to 100 mLSW2 concentrate).", "[exDNA extraction] Phosphate buffer prepa...
28,884
PCR Amplification of Desired Gene
null
dx.doi.org/10.17504/protocols.io.8fuhtnw
null
iGEM Dusseldorf
TITLE: PCR Amplification of Desired Gene AUTHORS: iGEM Dusseldorf [STEPS] ?. Pipette PCR components in the following order. Always set up at least two replicates. ?. ABCD1# Reactions[µl]Thermocycler ConditionsRounds2Water32.51. 98°C for 0:30 min3Green HF Buffer10.02. 98°C for 0:10 min5x4dNTPs (10 mM)1.03...
["Pipette PCR components in the following order. Always set up at least two replicates.", "ABCD1# Reactions[µl]Thermocycler ConditionsRounds2Water32.51. 98°C for 0:30 min3Green HF Buffer10.02. 98°C for 0:10 min5x4dNTPs (10 mM)1.03. X°C for 0:20 min5F Primer (10 µM)2.54. 72°C for X min6R Pri...
47,284
Direct cDNA Sequencing (SQK-DCS109)
4
dx.doi.org/10.17504/protocols.io.yxmvmkpxng3p/v1
https://www.protocols.io/view/direct-cdna-sequencing-sqk-dcs109-bseunbew
e.gustavsson
TITLE: Direct cDNA Sequencing (SQK-DCS109) AUTHORS: e.gustavsson [DESCRIPTION] This protocol describes how to prepare direct cDNA Sequencing libraries for nanopore sequencing without using PCR. [STEPS] SECTION: Reverse transcription and strand-switching 1. SECTION: Reverse transcription and strand-switching 2. Pr...
["[Reverse transcription and strand-switching]", "[Reverse transcription and strand-switching] Prepare the RNA in Nuclease-free water.", "[Reverse transcription and strand-switching] Transfer 100 ng into a 1.5 ml Eppendorf DNA LoBind tube.", "[Reverse transcription and strand-switching] Adjust the volume to up to 7.5 µ...
101,159
Isolation and Characterization of Tissue and Cell-Derived Extracellular Vesicles and Non-Vesicular Extracellular Particles
0
dx.doi.org/10.17504/protocols.io.5jyl822k8l2w/v2
https://www.protocols.io/view/isolation-and-characterization-of-tissue-and-cell-de2f3gbn
Marta Garcia Contreras, Emeli Chatterjee, Abhik Chakraborty, Michail Spanos, Kriti Bomb, Priyanka Gokulnath, Jeffery L. Franklin, Erika Duggan, John P. Nolan, Priyadarshini Pantham, Louise C. Laurent, Saumya Das
TITLE: Isolation and Characterization of Tissue and Cell-Derived Extracellular Vesicles and Non-Vesicular Extracellular Particles AUTHORS: Marta Garcia Contreras, Emeli Chatterjee, Abhik Chakraborty, Michail Spanos, Kriti Bomb, Priyanka Gokulnath, Jeffery L. Franklin, Erika Duggan, John P. Nolan, Priyadarshini Pantham,...
["[Processing of Conditioned Culture Media following Tissue and Cell Culture] Culture DiFi cells11 under the appropriate culture conditions as previously\npublished. BeWo cells were cultured as published earlier12,13 with the\nmodification of addition of 10% exosome-depleted FBS).", "[Processing of Conditioned Culture ...
68,580
Transforming Yeast (Instructor Protocol)
4
null
https://www.protocols.io/view/transforming-yeast-instructor-protocol-ce8cthsw
Brian Teague
TITLE: Transforming Yeast (Instructor Protocol) AUTHORS: Brian Teague [DESCRIPTION] This is the instructor protocol for The yeast transformation is adapted from Geitz and Schiestl: [GUIDELINES] We use dropout media components from Sunrise Science, but I'm sure media from other vendors is fine. Double-check,...
["[Prepare 250 ml of 2xYPD media] Put 250 mL of deionized water in a 250 ml or 500 ml bottle, then add:\n10 g \n5 g", "[Prepare 250 ml of 2xYPD media] Autoclave at 121 °C for 30 min on a liquid cycle.", "[Prepare 250 ml of 2xYPD media] Cool, then using good sterile technique, add 25 mLof a 40 Mass / % volume solution...