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null
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dx.doi.org/10.17504/protocols.io.nhedb3e
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
70,865
Protocol
1
dx.doi.org/10.17504/protocols.io.3byl4jke8lo5/v1
https://www.protocols.io/view/protocol-chfrt3m6
Tea Borkowska, Nikoloz Chkhartishvili, Ekaterine Karkashadze, Otar Chokoshvili, Pati gabunia, Lali Sharvadze, Tengiz Tsertsvadze
TITLE: Protocol AUTHORS: Tea Borkowska, Nikoloz Chkhartishvili, Ekaterine Karkashadze, Otar Chokoshvili, Pati gabunia, Lali Sharvadze, Tengiz Tsertsvadze [DESCRIPTION] Abstract Background Life expectancy and quality of life of people living with HIV have been dramatically improved after introducing antiretroviral ther...
["Prepare database just like one in the Materials section.", "Analyse the data contained in database.", "Formulate conclusions."]
60,154
2022 GenomeTrakr Proficiency Testing exercise (PulseNet Harmonized)
1
dx.doi.org/10.17504/protocols.io.e6nvw5m9dvmk/v1
https://www.protocols.io/view/2022-genometrakr-proficiency-testing-exercise-puls-b6y2rfye
Maria Balkey, Ruth Timme, Julie Haendiges
TITLE: 2022 GenomeTrakr Proficiency Testing exercise (PulseNet Harmonized) AUTHORS: Maria Balkey, Ruth Timme, Julie Haendiges [DESCRIPTION] This SOP outlines guidelines on how to process the isolates for the 2022 GenomeTrakr (GT) Proficiency Testing exercise. This SOP is applicable to all GenomeTrakr labs partici...
["[Culture Preparation] Salmonella and Escherichia/Shigella Lyophilized cultures:\n\nDay 1\n\nDocument the isolate number(s) and the lyophilized date(s) for your records. Wipe the aluminum cover and outside of the vial with isopropyl alcohol. Using sturdy forceps, aseptically remove the aluminum cover and rubber stoppe...
98,107
Ex vivo cell isolation
0
dx.doi.org/10.17504/protocols.io.q26g71b1kgwz/v1
https://www.protocols.io/view/ex-vivo-cell-isolation-db232qgn
Dorien De Pooter, Ben De Clerck, Koen Dockx, Domenica De Santis, Sarah Sauviller, Pascale Dehertogh, Matthias Beyens, Isabelle Bergiers, Isabel Nájera, Ellen Van Gulck, Nádia Conceição-Neto, Wim Pierson
TITLE: Ex vivo cell isolation AUTHORS: Dorien De Pooter, Ben De Clerck, Koen Dockx, Domenica De Santis, Sarah Sauviller, Pascale Dehertogh, Matthias Beyens, Isabelle Bergiers, Isabel Nájera, Ellen Van Gulck, Nádia Conceição-Neto, Wim Pierson [DESCRIPTION] This protocol details ex-vivo cell isolation. [STEPS] SECTION:...
["[Reagent preparation:] PBS-2% FCS: Thaw an aliquot of filtered FCS and prepare a solution of 2% FCS (vol/vol) in 1x PBS.", "[Reagent preparation:] 33.75% Percoll gradient: Prepare an isotonic solution of 33.75% Percoll using 10x PBS and 1x PBS-2% FCS. Prepare this solution fresh and store atRoom temperature and prote...
85,637
Human urine procurement and processing
1
dx.doi.org/10.17504/protocols.io.dm6gp3bmjvzp/v1
https://www.protocols.click/view/human-urine-procurement-and-processing-cxvdxn26
hannah.anvari, asma.giornazi, shriya.shah, maryellen.pavone, francesca.e.duncan francesca.duncan
TITLE: Human urine procurement and processing AUTHORS: hannah.anvari, asma.giornazi, shriya.shah, maryellen.pavone, francesca.e.duncan francesca.duncan [DESCRIPTION] Purpose: This protocol is intended for use in the collection and storage of human urine in a research setting. The protocol details the collection, proce...
["A urine sample is collected in a standard specimen cup during pre-op from a nurse. Once the urine sample has been collected it is placed immediately on ice following collection. A research coordinator will bring the urine sample to the lab on ice (2-4 °C). Urine samples should be transported in a blue, biohazard labe...
58,104
Pluripotency markers staining
4
dx.doi.org/10.17504/protocols.io.b4yyqxxw
https://www.protocols.io/view/pluripotency-markers-staining-b4yyqxxw
Hanqin Li, Dirk Hockemeyer, Frank Soldner
TITLE: Pluripotency markers staining AUTHORS: Hanqin Li, Dirk Hockemeyer, Frank Soldner [DESCRIPTION] This protocol describes the standard procedure for staining pluripotency markers, e.g. OCT4, SSEA4, alkaline phosphatase, and etc. on human pluripotent stem cells (hPSCs). Protocol overview A. Immunofluorescence st...
["[A. Immunofluorescence staining] Wash cells once with PBS", "[A. Immunofluorescence staining] Fix cells with 4% PFA at 4 Room temperature for 15 min", "[A. Immunofluorescence staining] Wash cells twice with PBS, incubate 5 min in between washes at Room temperature", "[A. Immunofluorescence staining] Incubate in 3% B...
null
null
null
dx.doi.org/10.17504/protocols.io.frjbm4n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Cas9 nuclease may be used <em>in vivo</em> to create targeted genome modifications.  There are several ways in which to introduce Cas9-guide RNA complexes into cells.  Here we present a method for the transfection of Cas9 RNP’s into HEK293 FT cells using Thermo Fisher Lipofec...
[]
91,481
Imaging axonal calcium dynamics in ex vivo mouse brain slices
1
dx.doi.org/10.17504/protocols.io.81wgbx9eolpk/v1
https://www.protocols.io/view/imaging-axonal-calcium-dynamics-in-ex-vivo-mouse-b-c5jzy4p6
Yan-Feng Zhang, Stephanie J Cragg
TITLE: Imaging axonal calcium dynamics in ex vivo mouse brain slices AUTHORS: Yan-Feng Zhang, Stephanie J Cragg [DESCRIPTION] This protocol describes how to image calcium dynamics in striatal dopaminergic axons in ex vivo mouse brain slices. We imaged calcium transients in response to single and trains (4 pulses, 100 ...
["[Image Acquisition] Change the exposure time to reach a frame rate of around 16.6 Hz every 2.5 min using Micro-Manager v1.4. 16.6 Hz frame rate every 2.5 min using Micro-Manager 1.4.", "[Image Analysis] The following steps were performed in MATLAB vR2019b and Fiji v1.5.\n\nExtract fluorescence intensity from the regio...
51,742
Isolation of Nucleated Cells from Whole Blood
1
dx.doi.org/10.17504/protocols.io.bwr6pd9e
https://www.protocols.io/view/isolation-of-nucleated-cells-from-whole-blood-bwr6pd9e
Steven B. Wells, Peter A. Szabo, Nora Lam
TITLE: Isolation of Nucleated Cells from Whole Blood AUTHORS: Steven B. Wells, Peter A. Szabo, Nora Lam [DESCRIPTION] This protocol describes a method for the isolation of pan-lymphocytes and pan-myeloid cells from human whole blood. By providing defined media formulations, volumes at each step, and a defined dilution...
["[Preparing Mediums and Buffers] Create the following IMDM-FBS-PSQ Media in a 500mL bottle of IMDM by using the table below: \n Component Volume (mL) Starting Conc.  Final Conc.* IMDM 500 - - Penicillin-Streptomycin-Glutamine 5 100X 1X FBS 50 100% 10%", "[Preparing Mediums and Buffers] Create...
102,454
Crystallisation of SARS-CoV-2 Mpro
1
dx.doi.org/10.17504/protocols.io.261ge5emyg47/v1
https://www.protocols.io/view/crystallisation-of-sars-cov-2-mpro-dgaw3sfe
blake.h.balcomb, Peter Marples, Lizbé Koekemoer, Daren Fearon, Charlie Tomlinson
TITLE: Crystallisation of SARS-CoV-2 Mpro AUTHORS: blake.h.balcomb, Peter Marples, Lizbé Koekemoer, Daren Fearon, Charlie Tomlinson [DESCRIPTION] The COVID-19 pandemic has highlighted the need to identify novel therapeutic interventions and strategies for pandemic preparedness against Severe Acute Respiratory Syndrome...
["[Crystallisation experiment] Protein and buffer requirements:\n43.2 µL5 mg/mL \n1.92 mL \n14.4 µL seeds, dilution 1:250", "[Crystallisation experiment] Dispense 20 µL into SwissCI 3 lens plate reservoir wells using a 100 µl multi-channel pipette.\nDispense 150 nL5 mg/mL to each lens using the SPT mosquito.\nDis...
28,855
Dechorionation of zebrafish embryos with Pronase for metronidazole-mediated ß-cell ablation
null
dx.doi.org/10.17504/protocols.io.8exhtfn
null
Sandra Rieger
TITLE: Dechorionation of zebrafish embryos with Pronase for metronidazole-mediated ß-cell ablation AUTHORS: Sandra Rieger [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block"><span>Zebrafish embryos develop in their eggshel...
["[Potential Pitfalls:]\n1. The embryos do not survive after treatment: This could be if embryos were incubated too long in Pronase. Decrease the incubation time and increase the number of washes.2. Dechorionation does not work: Pronase might be too old or animals are older than 24 hours post fertilization. Increase in...
33,918
Untitled
1
dx.doi.org/10.17504/protocols.io.bdc6i2ze
https://www.protocols.io/view/untitled-bdc6i2ze
Cui Jian
TITLE: Untitled AUTHORS: Cui Jian [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">l;l;l;</div></div> [STEPS] ?. [1] 0 µl 0 µl 0 µl ?. [1] {"blocks":[{"key":"8v8id","text":"The growing of microbrewerys in Brazil has imposing an increase in the production of different supplies for brewing, iincluding...
["[1]\n0 µl\n0 µl\n0 µl", "[1]\n{\"blocks\":[{\"key\":\"8v8id\",\"text\":\"The growing of microbrewerys in Brazil has imposing an increase in the production of different supplies for brewing, iincluding yeast biomass for beer fermentation. In Brazil,only four companies produce and sell yeasts for microbreweries, and on...
60,070
Mouse lung MAIT cell expansion and purification protocol
4
dx.doi.org/10.17504/protocols.io.x54v9yrpqg3e/v1
https://www.protocols.io/view/mouse-lung-mait-cell-expansion-and-purification-pr-b6werfbe
Gabriel A Ascui, Eleni Phung, Alba Mendis, Mitchell Kronenberg
TITLE: Mouse lung MAIT cell expansion and purification protocol AUTHORS: Gabriel A Ascui, Eleni Phung, Alba Mendis, Mitchell Kronenberg [DESCRIPTION] Mucosal Associated Invariant T (MAIT) cells are unconventional T cells present abundantly in human tissues. MAIT cells interact with antigens presented by MR1, an MHC cl...
["[MAIT cell in vivo expansion with BRD509] Salmonella enterica serotype Typhimurium BRD509 vaccine strain culture", "[MAIT cell in vivo expansion with BRD509] Day 0: Overnight culture of Salmonella Typhimurium BRD509 strain.\n\nPrepare 5 mL of LB media with 100 µg/ml of Streptomycin. \nPick -80 °C BRD509 stock and ino...
79,519
Prime Editing in Physcomitrium patens
4
dx.doi.org/10.17504/protocols.io.4r3l27r5qg1y/v1
https://www.protocols.click/view/prime-editing-in-physcomitrium-patens-crv7v69n
Pierre-François Perroud, Anouchka Guyon-Debast , Josep M Casacuberta, Wyatt Paul, Jean-Philippe Pichon, David Comeau, Fabien Nogué
TITLE: Prime Editing in Physcomitrium patens AUTHORS: Pierre-François Perroud, Anouchka Guyon-Debast , Josep M Casacuberta, Wyatt Paul, Jean-Philippe Pichon, David Comeau, Fabien Nogué [DESCRIPTION] PEG-mediated naked DNA protoplast transfection has been the standard transformation approach for more than thirty years...
["Using a binocular lens, check carefully the starting P. patens material for an eventual contamination (bacteria, fungus). The tissue must be green. An axenic culture is essential before performing any further experimental step.", "Let the protoplasts grow for 6 to 7 days on the transformation plates. Observe the plat...
68,928
Pole Test
1
dx.doi.org/10.17504/protocols.io.5jyl8938rv2w/v1
https://www.protocols.io/view/pole-test-cfi8tkhw
Sabina Marciano, Tae-In Kam, Roberta Marongiu, Ted Dawson
TITLE: Pole Test AUTHORS: Sabina Marciano, Tae-In Kam, Roberta Marongiu, Ted Dawson [DESCRIPTION] This behavior test is used to assess motor coordination in mice. [GUIDELINES] Main outcome measures: average time to turn, average time to reach the bottom [STEPS] 1. Place the mouse on the vertical pole with their head...
["Place the mouse on the vertical pole with their head oriented upwards 7.5cm from the top of the pole.", "During day 1, gently guide the mouse to turn around and go to the bottom of the pole. They are trained 3 times.", "On day 2, they are trained 3 times again without guidance.", "On day 3, the test day, mice are aga...
18,732
GUIDE-seq simplified library preparation protocol (CRISPR/Cas9 off-target cleavage detection)
null
dx.doi.org/10.17504/protocols.io.wikfccw
null
Nagendra palani
TITLE: GUIDE-seq simplified library preparation protocol (CRISPR/Cas9 off-target cleavage detection) AUTHORS: Nagendra palani [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">GUIDE-seq is an experimental method to detect off-target cleavages caused during CRISPR/Cas editing.</div><div class = "text-b...
["[Adaptor formation]\nUse 0.1x TE to prepare 100 μM stock of all the oligos.In a 0.2 ml PCR tube, add the following reagents, vortex well, and spin down. Place on the thermal cycler at for (heated lid). -------------------------------------------- After incubation, terminate the incubation and let the heat block...
61,943
Bioconjugation Strategies
1
null
https://www.protocols.io/view/bioconjugation-strategies-b8qxrvxn
BOC Sciences
TITLE: Bioconjugation Strategies AUTHORS: BOC Sciences [DESCRIPTION] Bioconjugation is a chemical technique that is used to couple two molecules together, at least one of which is a biomolecule, such as carbohydrate, nucleic acid, or protein. Functionalization of polymers can provide improved stability, solubilit...
[]
26,146
What is CD14
null
dx.doi.org/10.17504/protocols.io.5sag6ae
null
susan wind
TITLE: What is CD14 AUTHORS: susan wind [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">CD14, the LPS (Lipopolysaccharide) receptor, was originally a leukocyte differentiation antigen present on the surface of cells such as monocytes and macrophages. It was first discovered by TODD from the surface ...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.q83dzyn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>Fluorescent staining for Callose of sieve tubes of phloem.</strong></p> <p>Calose (β-1→3-glucan) is the distinguished polysaccharide present on the sieve plates, of each sieve tube member of phloem tissue.  Aniline blue stain specifically callose and it is use as a ma...
[]
41,461
SUPER rapid kit
4
dx.doi.org/10.17504/protocols.io.bkqvkvw6
https://www.protocols.io/view/super-rapid-kit-bkqvkvw6
Hyeon Jin Kim, Seong T. Hong
TITLE: SUPER rapid kit AUTHORS: Hyeon Jin Kim, Seong T. Hong [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">We developed a supersensitive capillary reaction kit intended for the direct and qualitative detection of SARS-CoV-2 virus in saliva and nasal swabs. It enables sensitive and specific detecti...
["Prepare the specimen by collecting saliva, sputum, or nasal swab for testing", "Dispense 2~5 drops of the specimen into the sampling tube (provided) filled with a general transfer pipette", "Mix the content by pipetting up & down gently", "Dispense 5 drops of mixed samples into the sample well of the cassette (provid...
60,170
Golden Gate Assembly
4
dx.doi.org/10.17504/protocols.io.x54v9yr3mg3e/v1
https://www.protocols.io/view/golden-gate-assembly-b6zirf4e
Isaac Núñez, Tamara Matute, Fernan Federici
TITLE: Golden Gate Assembly AUTHORS: Isaac Núñez, Tamara Matute, Fernan Federici [DESCRIPTION] The Golden Gate technique allows the assembly of genetic sequences from libraries of standardized basic components, which are cleaved from their donor vectors and concatenated in the acceptor vector in a defined order. Th...
["[Preparation of the DNA components] Prepare plasmids stock solutions of the desired components to be used.", "[Preparation of the DNA components] Measure the concentration of purified plasmids.", "[Preparation of the DNA components] Perform dilutions of the plasmids to working concentrations. \nIt is:\n15 fmol/μL for...
82,281
Eastern Hemlock Tissue Collection for DNA
4
null
https://www.protocols.click/view/eastern-hemlock-tissue-collection-for-dna-cukhwut6
Karl Fetter
TITLE: Eastern Hemlock Tissue Collection for DNA AUTHORS: Karl Fetter [DESCRIPTION] Steps for collecting tissue from Eastern Hemlocks for DNA analysis. The Plant Computational Genomics lab is conducting a landscape genomics study of climate adaptation in the Eastern Hemlock. This protocol is a step-by-step guide to co...
["[Introduction] This protocol is intended for field collection of leaf tissue for the Eastern Hemlock conservation genomics project sponsored by the Plant Computation Genomics lab at the University of Connecticut. The goal of the project is to identify climate adapted genomic variation for seed banking and potential u...
96,260
REDI-NET F-1 FECES FIELD SAMPLING
0
dx.doi.org/10.17504/protocols.io.yxmvm3dx6l3p/v1
https://www.protocols.io/view/redi-net-f-1-feces-field-sampling-c99cz92w
REDI-NET Consortium
TITLE: REDI-NET F-1 FECES FIELD SAMPLING AUTHORS: REDI-NET Consortium [DESCRIPTION] To outline steps for properly collecting fecal samples from pastures to evaluate the risk of zoonotic disease transmission by the detection of pathogens from vertebrate DNA (vDNA). [BEFORE_START] Know the risks associated with the stu...
["[SAMPLE SITE SELECTION] Locate a fecal sample and estimate its age in days and score its consistency.", "[SAMPLE SITE SELECTION] Age Estimate", "[SAMPLE SITE SELECTION] Consistency Score - Cattle (see photos in Appendix 1)", "[SAMPLE SITE SELECTION] Consistency Score - Sheep (see photos in Appendix 2)", "[SAMPLE SITE...
60,297
Nuclei isolation from frozen human brain samples for snRNA-seq
4
dx.doi.org/10.17504/protocols.io.j8nlkk9ydl5r/v1
https://www.protocols.io/view/nuclei-isolation-from-frozen-human-brain-samples-f-b65hrg36
Marcos Nascimento
TITLE: Nuclei isolation from frozen human brain samples for snRNA-seq AUTHORS: Marcos Nascimento [DESCRIPTION] Protocol to isolate nuclei from snap-frozen human brain samples for sn-RNAseq [STEPS] SECTION: Preparation 2. Clean your work area (bench and pipettes) with RNAse Zap. SECTION: Nuclei Isolation 6. Transfer t...
["[Preparation] Clean your work area (bench and pipettes) with RNAse Zap.", "[Nuclei Isolation] Transfer the LB-sample mix to a labelled glass dounce homogenizer . Add 2ml of LB, bringing the total volume in the douncer to 3mL.on ice \nYou can add 1ml of LB to the sample eppendorf to collect any remaining tissue.", "[N...
null
null
null
dx.doi.org/10.17504/protocols.io.hd5b286
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
98,447
CODA: shorthand for calling functions | HuBMAP | JHU-TMC
0
dx.doi.org/10.17504/protocols.io.5qpvoknkbl4o/v2
https://www.protocols.io/view/coda-shorthand-for-calling-functions-hubmap-jhu-tm-dcdp2s5n
Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Sashank Reddy, Denis Wirtz
TITLE: CODA: shorthand for calling functions | HuBMAP | JHU-TMC AUTHORS: Kyu Sang Han, Pei-Hsun Wu, Joel Sunshine, Ashley Kiemen, Sashank Reddy, Denis Wirtz [DESCRIPTION] To downsample ndpi or svs images to 10x, 5x, and 1x tifs, use this function: create_downsampled_tif_images or try Openslide in pytho...
["[shorthand in the abstract] Use the above shorthand to facilitate your workflow by using it as a \"cheat sheet\""]
null
null
null
dx.doi.org/10.17504/protocols.io.c9iz4d
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol combines phage gene detection with rRNA detection for the identification of host cells and detection of free phage particles. [BEFORE_START] <strong>Before staring the phageFISH experiment: Probe Design, Probe Synthesis, and Hybridization Stringency:</strong><br /...
[]
25,576
Expression and purification of (GST-tagged) (Kai) proteins
null
dx.doi.org/10.17504/protocols.io.48ggztw
null
Anika Wiegard, Christin Köbler, Katsuaki Oyama, Anja K. Dörrich, Chihiro Azai, Kazuki Terauchi, Annegret Wilde, Ilka Maria Axmann
TITLE: Expression and purification of (GST-tagged) (Kai) proteins AUTHORS: Anika Wiegard, Christin Köbler, Katsuaki Oyama, Anja K. Dörrich, Chihiro Azai, Kazuki Terauchi, Annegret Wilde, Ilka Maria Axmann [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol can be used for:</div><div class ...
["[heterologous protein expression in E.coli]\ntransformation:", "[heterologous protein expression in E.coli]\npre-culture:inoculate 100 ml terrific broth medium containing 100 µg ampicillin ml-1 with resulting transformantsincubate over night at 37 °C and 200-250 r.p.m.", "[heterologous protein expression in E.coli]\n...
23,774
RNA extractions from de-etiolated Arabidopsis seedlings using CTAB
null
dx.doi.org/10.17504/protocols.io.3f6gjre
null
Akila Wijerathna-Yapa, Andrew Bowerman, Diep Ganguly
TITLE: RNA extractions from de-etiolated Arabidopsis seedlings using CTAB AUTHORS: Akila Wijerathna-Yapa, Andrew Bowerman, Diep Ganguly [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A CTAB based method for extracting RNA, which is particularly useful for tough tissues. This has been adapted from a...
["Harvest tissues into liquid nitrogen and grind (under liquid nitrogen) using mortar and pestle to obtain a fine powder. Ground tissue should be kept in safe-lock tubes in liquid nitrogen (or returned to -80C storage) until all samples are processed. This step is critical for efficient extractions so take your time he...
22,315
Universal DNA isolation protocol
null
dx.doi.org/10.17504/protocols.io.z2jf8cn
null
Ruslan Kalendar
TITLE: Universal DNA isolation protocol AUTHORS: Ruslan Kalendar [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The isolation of nucleic acids from a sample is an important step for many molecular biological applications and medical diagnostic assays. This protocol describes an efficient method for...
["In 2 ml tube with mechanically disrupted seeds/leaves/herbarium or DNA solution (CTAB purification) add 1 ml CTAB solution buffer with RNAse A (the sample mass should not exceed 100 mg), vortex very well and incubate the samples at 60-65°C during 60-120 min or longer (long incubation increases DNA yield).", "Centrifu...
28,402
MojoSort™ Mouse NK Cell Isolation Kit Protocol
null
dx.doi.org/10.17504/protocols.io.7yshpwe
null
Sam Li
TITLE: MojoSort™ Mouse NK Cell Isolation Kit Protocol AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Product description and procedure summary:</span><span> Target cells are depleted by incubating your sample with the biotin antibody cocktail follow...
["Prepare cells from your tissue of interest or blood without lysing erythrocytes.", "In the final wash of your sample preparation, resuspend the cells in MojoSort™ Buffer by adding up to 4 mL in a 5 mL (12 x 75 mm) polypropylene tube.Note: Keep MojoSort™ Buffer on ice throughout the procedure.", "Filter the cells with...
27,559
Human embryonic gonad dissociation with Trypsin-EDTA
1
dx.doi.org/10.17504/protocols.io.66fhhbn
https://www.protocols.io/view/human-embryonic-gonad-dissociation-with-trypsin-ed-66fhhbn
Regina Hoo, Roser Vento-Tormo, Carmen Sancho
TITLE: Human embryonic gonad dissociation with Trypsin-EDTA AUTHORS: Regina Hoo, Roser Vento-Tormo, Carmen Sancho [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for enrichment of fetal gonadal cells</div></div> [STEPS] ?. Samples arrive in hypothermosol solution. The gonad is wash...
["Samples arrive in hypothermosol solution. The gonad is washed with PBS and dissociated into single-cells following the protocol below.", "Fragment the gonads with a scalpel into 2 to 20 pieces per gonad (depending on developmental stage).", "Suspend gonadal fragments into a 15 mL tube containing of Trypsin-EDTA (0.25...
94,180
Barnes Maze Protocol
4
dx.doi.org/10.17504/protocols.io.kxygx3bozg8j/v1
https://www.protocols.io/view/barnes-maze-protocol-c78czrsw
Lisa Blackmer-Raynolds, Ian N Krout, Tim Sampson
TITLE: Barnes Maze Protocol AUTHORS: Lisa Blackmer-Raynolds, Ian N Krout, Tim Sampson [DESCRIPTION] The Barnes Maze test is a test of rodent’s spatial learning and memory abilities. In this test, animals are placed in an aversive environment (with a loud noise and bright lights) and trained to locate a hidden escape b...
["[Set-up] Set up extra bright overhead lamps directly above maze so there is no difference in the brightness across the maze.", "[Set-up] Set white noise maker to highest volume setting (butkeep off for now).", "[Set-up] Make sure there are landmarks around the room that the animals can use to orient, everything in th...
27,910
private parent
null
dx.doi.org/10.17504/protocols.io.7hehj3e
null
Monica Hassan
TITLE: private parent AUTHORS: Monica Hassan [STEPS] ?. music
["music"]
19,753
Bioelectrochemistry protocol For CHI Potentiostat
null
dx.doi.org/10.17504/protocols.io.xihfkb6
null
Rose Jones Jones, Beth Orcutt
TITLE: Bioelectrochemistry protocol For CHI Potentiostat AUTHORS: Rose Jones Jones, Beth Orcutt [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The following protocol describes how to prepare microbial fuel cells (MFCs) with Indium Tin Oxide (ITO) electrodes for use as a poised potential enrichment ...
["Preparing ITO “working” electrodes and carbon cloth “counter” electrodes Measure and cut 5 lengths of Ti wire 8cm long. Rub well with 100% ethanol and kimwipe to remove impurities (These will show up as black marks on the kimwipe. Measure and cut four 1.5” x 1.5” pieces of carbon cloth for the counter electrodes, an...
91,122
F-5 FECES SHIPPING
4
dx.doi.org/10.17504/protocols.io.x54v9pzy4g3e/v1
https://www.protocols.io/view/f-5-feces-shipping-c48syzwe
REDI-NET Consortium
TITLE: F-5 FECES SHIPPING AUTHORS: REDI-NET Consortium [DESCRIPTION] This protocol describes feces shipping. [GUIDELINES] OBJECTIVE To outline steps for proper packaging and shipping of preserved feces samples from a REDI-NET Silver Lab to a REDI-NET Gold Lab. SUMMARY/SCOPE The overarching aim of the REDI-NET is t...
["[SAMPLE PREPARATION] Primary holding sample holding containers must be leak-proof. Samples should be preserved in appropriate preservative, if applicable (REDI-NET Feces Processing SOP F-2). Ensure that the lids are tightly closed to prevent leaking of storage media while in transit.", "[SAMPLE PREPARATION] Pack vial...
36,002
Cell line information
1
dx.doi.org/10.17504/protocols.io.rm7vz82o8vx1/v1
https://www.protocols.io/view/cell-line-information-bfeajjae
Philippa R Kennedy
TITLE: Cell line information AUTHORS: Philippa R Kennedy [DESCRIPTION] An amalgamation of cell line data in the lab. Its origin and standard culture conditions. [STEPS] SECTION: Lung 3. Non-small cell lung cancers: Lung adenocarcinomas: 1. A549 (RRID:CVCL_0023) was purchased from the American Type Culture Collect...
["[Lung] Non-small cell lung cancers: \n\nLung adenocarcinomas: \n1. A549 (RRID:CVCL_0023) was purchased from the American Type Culture Collection (ATCC) in March 2019. \n2. NCI-H322 (RRID:CVCL_1556; purchased May 2019, Sigma Aldrich)\n3. NCI-H522 (RRID: CVCL_1567).\n\nLarge cell lung carcinoma: \n1. NCI-H460 (RRID:CVC...
null
null
null
dx.doi.org/10.17504/protocols.io.dp55q5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] For use in "<a href="https://www.protocols.io/view/Isolation-of-cyanophages-by-liquid-bioassays-dp25qd" target="_blank">Isolation of cyanophages by liquid bioassays</a>" [STEPS] ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.paudiew
null
null
TITLE: No Title AUTHORS: [STEPS] ?.
[]
95,842
Elutriator operation protocol
0
dx.doi.org/10.17504/protocols.io.36wgq3x6olk5/v1
https://www.protocols.io/view/elutriator-operation-protocol-c9uaz6se
Casey A Schlenker
TITLE: Elutriator operation protocol AUTHORS: Casey A Schlenker [DESCRIPTION] Operation of The OSU Soybean Pathology and Nematology elutriator [STEPS] SECTION: Turning elutriator on 1. Turn on the water supply by turning the red valve on the wall counterclockwise. Open the flow for both hoses by turning the small yel...
["[Turning elutriator on] Turn on the water supply by turning the red valve on the wall counterclockwise. Open the flow for both hoses by turning the small yellow handles parallel to the hoses.", "[Turning elutriator on] Turn on the air compressor, located outside the greenhouse door behind the metal gate", "[Turning e...
54,254
Mercury Sequence and Sample Metadata Prep for Submission Workflow on the Terra Platform
5
null
https://www.protocols.io/view/mercury-sequence-and-sample-metadata-prep-for-subm-by8npzve
Jill V Hagey, Kevin Libuit, Lingzi Xiaoli, Technical Outreach and Assistance for States Team
TITLE: Mercury Sequence and Sample Metadata Prep for Submission Workflow on the Terra Platform AUTHORS: Jill V Hagey, Kevin Libuit, Lingzi Xiaoli, Technical Outreach and Assistance for States Team [DESCRIPTION] Public health laboratories are encouraged to submit sequencing data for SARS-CoV-2 to multiple public dat...
["[Where to Begin] This workflow is intended for users of Titan workflows for SARS-CoV-2 strain characterization on the Terra platform and expects the following protocols have already been completed: \n\nTerra and Google Cloud Accounts\nSamples uploaded\nOutput from the Titan workflow\n\nThis protocol is intended only ...
58,874
Beam Break and Nesting
4
dx.doi.org/10.17504/protocols.io.b5q2q5ye
https://www.protocols.io/view/beam-break-and-nesting-b5q2q5ye
Haley Geertsma
TITLE: Beam Break and Nesting AUTHORS: Haley Geertsma [DESCRIPTION] This protocol is used to test mice in the Beam Break and Nesting behavioural test. [STEPS] 2. Single-cage mice in the Beam Break rack with enough food and water for 48-hours. 1. Start Fusion software to begin recording and confirm the lights are on...
["Single-cage mice in the Beam Break rack with enough food and water for 48-hours.", "Start Fusion software to begin recording and confirm the lights are on the appropriate light-dark cycle.", "Turn off Fusion software and add a square of cotton nestlet. Leave mice for 17-19 hours.", "Record and/or score nests as per h...
71,672
Growing Arabidopsis on nutrient-rich media
4
dx.doi.org/10.17504/protocols.io.bp2l6917dlqe/v1
https://www.protocols.io/view/growing-arabidopsis-on-nutrient-rich-media-ch8yt9xw
Diep R Ganguly
TITLE: Growing Arabidopsis on nutrient-rich media AUTHORS: Diep R Ganguly [DESCRIPTION] Notes on growing Arabidopsis seedlings on nutrient-rich media in plates. [STEPS] SECTION: Prepare media 1. Select and prepare autoclaved media of choice (see recipes below). Make sure to pH before adding gelling agent. MS basa...
["[Prepare media] Select and prepare autoclaved media of choice (see recipes below). Make sure to pH before adding gelling agent. \n\n MS basal salts\t 2.15 – 4.33 g/LSucrose10-30 g/LVitamins (1000X) (optional)1 mL/L\tMES (optional)0.1% (w/v)KOHpH 5.75Agar/Phytoblend8 g/L\n\n Gamborg’s B-5 basal medium3.21 g/LSucrose...
26,842
New Iron Extracting Method from Cattle's Blood for Iron Concentration Analysis
null
dx.doi.org/10.17504/protocols.io.6f2hbqe
null
Khairil Asnan, Mohd. Sadzan Asram bin Aliming, Aprialdy Idrus, Diana Eka Pratiwi
TITLE: New Iron Extracting Method from Cattle's Blood for Iron Concentration Analysis AUTHORS: Khairil Asnan, Mohd. Sadzan Asram bin Aliming, Aprialdy Idrus, Diana Eka Pratiwi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:justify"><span>Cattle's blood com...
["[Sample preparation]\nCompress the whole blood sample from cattle.\n6 ml\n37 °C", "[Sample preparation]\nMix sample with\n[blood sample]\n[NaOH]\n37 °C\ndark green solution with strong odor", "Chelate reaction by adding into the previous mixed sample solution then let it sit for . After that, mixed them by for .\...
21,888
RNAi by feeding in Euplotes focardii (povisional)
null
dx.doi.org/10.17504/protocols.io.zk8f4zw
null
Francesca Papi, Angela Piersanti
TITLE: RNAi by feeding in Euplotes focardii (povisional) AUTHORS: Francesca Papi, Angela Piersanti [STEPS] ?. Grow RNAse III deficient E.coli strain HT115 in LB with antibiotic selection o/n at 37°C. ?. Prepare a 1:100 dilution of the bacterial culture, and grow it at 37°C until it reaches an OD600 of 0.4. ?. Add 0.4 ...
["Grow RNAse III deficient E.coli strain HT115 in LB with antibiotic selection o/n at 37°C.", "Prepare a 1:100 dilution of the bacterial culture, and grow it at 37°C until it reaches an OD600 of 0.4.", "Add 0.4 mM IPTG, and induce RNA transcription from the L4440 plasmid, in which it is included the gene of interest t...
33,612
Case Processing SOP (Spleen)
null
dx.doi.org/10.17504/protocols.io.bc3kiykw
null
Marda Jorgensen
TITLE: Case Processing SOP (Spleen) AUTHORS: Marda Jorgensen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The purpose of this Standard Operating Procedure is to outline procedures for processing and storing spleen received for HuBMAP consortium assay and analysis. </div></div> [STEPS] ?. The tis...
["The tissues received will be identified as follows: Spleen (SP)", "Sample prepared from tissues will be identified using the following nomenclature, abbreviations, and formats: Formalin Fixed Paraffin Embedded---FFPE (paraffin block)Formalin Fixed Frozen Embedded---Fixed OCT BlockFresh Frozen Embedded---Fresh OCT Blo...
59,440
test protocol
4
null
https://www.protocols.io/view/test-protocol-b6aqradw
Julien J Ghislain
TITLE: test protocol AUTHORS: Julien J Ghislain [DESCRIPTION] xx [STEPS] 1. xxx 2. xxx
["xxx", "xxx"]
40,962
VISUALIZING PROTEINS ON PAGE GEL WITH COOMASSIE COLLOIDAL STAIN - CHEM 584
4
dx.doi.org/10.17504/protocols.io.bj9akr2e
https://www.protocols.io/view/visualizing-proteins-on-page-gel-with-coomassie-co-bj9akr2e
Ken Christensen
TITLE: VISUALIZING PROTEINS ON PAGE GEL WITH COOMASSIE COLLOIDAL STAIN - CHEM 584 AUTHORS: Ken Christensen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Staining polyacrylamide gels using a colloidal Coomassie Blue stain. Adapted from a JoVE article.</div></div> [STEPS] ?. Wash gel with ~50 mL dd...
["Wash gel with ~50 mL ddH2O 3x for ~ for each wash on a shaker NOTE: insufficient washing causes poor sensitivity because the remaining SDS on the gels disturbs the bounding of the dye to the protein", "Shake the colloidal Coomassie solution before use to disperse particles evenly.", "Incubate the gels with the Coomas...
90,064
Sectioning and HE staining of Mouse Brain and embryo by Cryostat
4
dx.doi.org/10.17504/protocols.io.5qpvo379dv4o/v1
https://www.protocols.io/view/sectioning-and-he-staining-of-mouse-brain-and-embr-c37qyrmw
Yuting Fu, jiashikai, Xiaodong Liu
TITLE: Sectioning and HE staining of Mouse Brain and embryo by Cryostat AUTHORS: Yuting Fu, jiashikai, Xiaodong Liu [DESCRIPTION] This protocol describes how to use the cryostat to prepare and slice mouse brain and embryo sections for HE staining and following spatial transcriptomic experiments. [STEPS] SECTION: Prep...
["[Preparation Methods for E12.5 mouse-embryo-eye] Mouse embryos were collected from pregnant C57BL/6J female mice at embryonic day 12.5\n(E12.5). Mouse brain was dissected from 8-week-old C57BL/6J male mice;", "[Preparation Methods for E12.5 mouse-embryo-eye] E12.5 pregnant female mice was anesthetized with carbon dio...
71,313
Extraction of Herbarium material for chemical profiling
1
null
https://www.protocols.io/view/extraction-of-herbarium-material-for-chemical-prof-chvrt656
Ricardo M. Borges
TITLE: Extraction of Herbarium material for chemical profiling AUTHORS: Ricardo M. Borges [DESCRIPTION] From the perspective of the documentation of plant biological diversity, herbaria are the repository of intraspecific and interspecific variability. The rich assortments contained in these collections essay the div...
["[Sample preparation] Organize every sample that will be used in each study into an EXCEL Sheet;\ninclude information and comments on every sample\nhave information about classification grouping on every sample\neach sample should also have the filename for their analytical data indexed (these can be added afterward)\...
42,390
Mobile Device Quantification of Lateral Flow Tests: Modular illumination and sensor chamber (US patent US20160131592A1)
3
dx.doi.org/10.17504/protocols.io.bmmwk47e
https://www.protocols.io/view/mobile-device-quantification-of-lateral-flow-tests-bmmwk47e
Donald Cooper
TITLE: Mobile Device Quantification of Lateral Flow Tests: Modular illumination and sensor chamber (US patent US20160131592A1) AUTHORS: Donald Cooper [STEPS]
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ea6bahe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol is part of the VERVE holiday drive to collect exotic off-the-shelf laboratory recipes. More details <a href="https://www.protocols.io/g/verve-net/news/call-for-recipes" target="_blank">here</a>. [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?...
[]
64,610
Via Keto Gummies Reviews UK: Is It Best Fat Burner
1
dx.doi.org/10.17504/protocols.io.6qpvr6w6ovmk/v1
https://www.protocols.io/view/via-keto-gummies-reviews-uk-is-it-best-fat-burner-cbcasise
viaketohack
TITLE: Via Keto Gummies Reviews UK: Is It Best Fat Burner AUTHORS: viaketohack [DESCRIPTION] These days, many individuals are attempting to keep up with solid weight and every one of these are expected to help them in having a sound life. In any case, getting in shape while having solid absorption and common heartbe...
[]
95,357
Standardised Methods for Feed Tabs Preparation to Assess Feeding Rate and Exposure to Insoluble Substances in Parhyale hawaiensis
4
dx.doi.org/10.17504/protocols.io.81wgbx3qylpk/v1
https://www.protocols.io/view/standardised-methods-for-feed-tabs-preparation-to-c9c5z2y6
Ibrahim Lawan, Dominique MJ Anderson, Theodore, B. Henry
TITLE: Standardised Methods for Feed Tabs Preparation to Assess Feeding Rate and Exposure to Insoluble Substances in Parhyale hawaiensis AUTHORS: Ibrahim Lawan, Dominique MJ Anderson, Theodore, B. Henry [DESCRIPTION] This protocol outlines standardised procedures for preparing feed tabs to assess feeding rates and con...
["[Introduction] Dietary exposure and toxicological assessments of insoluble substances represent emerging frontiers in aquatic ecotoxicology, necessitating the development of standardised bioassays that accurately reflect real-world exposure scenarios (Connon et al., 2012). Current toxicity testing methodologies prima...
null
null
null
dx.doi.org/10.17504/protocols.io.cpdvi5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Protocol for non-radioactive phosphorylation with T4 PNK. [STEPS] ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.fd3bi8n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Instructions to make 50 mL of incubation buffer for protoplast isolation.</p> <p>Adapted from Yoo et al. 2007 http://www.nature.com/nprot/journal/v2/n7/full/nprot.2007.199.html</p> [STEPS] ?. ?. ?.
[]
62,574
Canadian Airborne Biodiversity Observatory&#39;s Forest Inventory Field Survey Protocol
1
dx.doi.org/10.17504/protocols.io.q26g7rn23vwz/v2
https://www.protocols.io/view/canadian-airborne-biodiversity-observatory-39-s-fo-b9cnr2ve
Anna L Crofts, Sabine St-Jean, Mark Vellend
TITLE: Canadian Airborne Biodiversity Observatory&#39;s Forest Inventory Field Survey Protocol AUTHORS: Anna L Crofts, Sabine St-Jean, Mark Vellend [DESCRIPTION] Here, we describe the standardized protocol used by the Canadian Airborne Biodiversity Observatory (CABO) to conduct the field-based surveys of canopy tree...
["[Plot Establishment in the Field and in Fulcrum] Systematically establish plots to span the range of conditions across the imaged area. Specifically, we aim to:\n\n Maximize the number of unique tree species assemblages\n Cover the range of topographic conditions (e.g., elevation, slope, and aspect)", "[Obtai...
98,735
Patient PBMC collection and cryopreservation and cryorecovery
0
dx.doi.org/10.17504/protocols.io.n92ld895ov5b/v1
https://www.protocols.io/view/patient-pbmc-collection-and-cryopreservation-and-c-dcnp2vdn
Rebecca Wallings
TITLE: Patient PBMC collection and cryopreservation and cryorecovery AUTHORS: Rebecca Wallings [DESCRIPTION] Isolation of PBMC from human blood using CPT tubes, cryopreservation and cryorecovery of cells [STEPS] SECTION: PBMC collection and cryopreservation 1. Collect 8 mL of blood per BD Vacutainer CPT Cell Preparat...
["[PBMC collection and cryopreservation] Collect 8 mL of blood per BD Vacutainer CPT Cell Preparation Tube with Sodium Citrate (BD Biosciences, 362761). \n\nInvert tubes 8-10 times and centrifuged at room temperature at 1500x g for 20 minutes with no brake. \n\nUsing a 10mL stripette , transfer the PBMC enriched layer ...
58,462
Magnitude of job satisfaction and intention to leave present job among nurses in selected Federal Hospitals in Addis Ababa, Ethiopiantitled collection
2
dx.doi.org/10.17504/protocols.io.b5b6q2re
https://www.protocols.io/view/magnitude-of-job-satisfaction-and-intention-to-lea-b5b6q2re
Aynye Negese*, Elsabet Getye2, ziyadahm
TITLE: Magnitude of job satisfaction and intention to leave present job among nurses in selected Federal Hospitals in Addis Ababa, Ethiopiantitled collection AUTHORS: Aynye Negese*, Elsabet Getye2, ziyadahm [DESCRIPTION] Background Job dissatisfaction issues and Health workers’ intention to leave is an increas...
[]
90,468
C-SOP-201: Genomic DNA Quantification using a Qubit Fluorometer
4
dx.doi.org/10.17504/protocols.io.kqdg3xb47g25/v1
https://www.protocols.io/view/c-sop-201-genomic-dna-quantification-using-a-qubit-c4kcyusw
Mihir Kekre, Ben Pascoe
TITLE: C-SOP-201: Genomic DNA Quantification using a Qubit Fluorometer AUTHORS: Mihir Kekre, Ben Pascoe [DESCRIPTION] In order to prepare high quality double-stranded DNA libraries for Illumina WGS, extracted genomic DNA needs to be accurately quantified. Qubit Fluorometers detect fluorescent dyes that are specific to...
["[Before starting] The buffer and BR/HS reagent (dye) should be stored at 4 Room temperature while the standards must be stored at 4 °C. \n\nThe reagent is light and humidity sensitive, it should be stored in the dark and exposed minimally during assay and sample preparation. When stored as directed, the reagent has a...
61,548
cDNA Library Preparation for scRNA-seq (10x Genomics)
5
dx.doi.org/10.17504/protocols.io.bp2l6175rvqe/v1
https://www.protocols.io/view/cdna-library-preparation-for-scrna-seq-10x-genomic-b8ckrsuw
molmer , Martin Lotz, Tony Mondala, Steven Head
TITLE: cDNA Library Preparation for scRNA-seq (10x Genomics) AUTHORS: molmer , Martin Lotz, Tony Mondala, Steven Head [DESCRIPTION] Samples are processed using V2 barcoding chemistry kits of 10x Genomics. For each run, 10,000 cells from individual donors are labeled with distinct oligo-barcoded antibodies (cell h...
["Single cells are isolated from human femoral articular cartilage using doi \ndx.doi.org/10.17504/protocols.io.14egn765qv5d/v1.", "Single cell suspensions are multiplexed and converted to barcoded scRNAseq libraries using the Chromium Single Cell 3’ Library, Gel Bead and Chip Kit (10x Genomics), and the BD™ Hu Single ...
63,402
Oprah Winfrey ACV Gummies Reviews: Shark tank gummy candy 2022
1
dx.doi.org/10.17504/protocols.io.rm7vzy2nrlx1/v1
https://www.protocols.io/view/oprah-winfrey-acv-gummies-reviews-shark-tank-gummy-b96ir9ce
oprahwinfreyreviewsads
TITLE: Oprah Winfrey ACV Gummies Reviews: Shark tank gummy candy 2022 AUTHORS: oprahwinfreyreviewsads [DESCRIPTION] Oprah Winfrey ACV Gummies Reviews: Shark tank gummy candy 2022 [STEPS] 1. Oprah Winfrey ACV Gummies Reviews: Shark tank gummy candy 2022 Oprah Winfrey Keto – The Supplement for a Spare Shape and Sli...
["Oprah Winfrey ACV Gummies Reviews: Shark tank gummy candy 2022\n\nOprah Winfrey Keto – The Supplement for a Spare Shape and Slim Body! \n\nThere'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 a...
93,935
Protocol: Purification of DNA from Whole Blood using the QIAamp Blood Midi Kit (Spin Protocol) + QC with Qubit fluorometer
6
dx.doi.org/10.17504/protocols.io.n92ldmx3ol5b/v1
https://www.protocols.io/view/protocol-purification-of-dna-from-whole-blood-usin-c7ypzpvn
Paula Saffie, Breeana Baker, Kimberley J Billingsley
TITLE: Protocol: Purification of DNA from Whole Blood using the QIAamp Blood Midi Kit (Spin Protocol) + QC with Qubit fluorometer AUTHORS: Paula Saffie, Breeana Baker, Kimberley J Billingsley [DESCRIPTION] The QIAamp DNA Blood Midi procedure yield pure DNA ready for direct restriction digestion or amplification. With ...
["[DNA extraction procedure]", "[Sample QC procedure (Qubit Fluorometer)] STANDARDIZE THE QUBIT (every time you use it or once a week according to lab protocol)", "[Sample QC procedure (Qubit Fluorometer)] Get0.2 mL Qubit tubes.", "[Sample QC procedure (Qubit Fluorometer)] In one strip, add10 mL of BR Standard 1.", "[S...
97,794
Visual frailty in the ageing population: A scoping review
0
dx.doi.org/10.17504/protocols.io.x54v92844l3e/v1
https://www.protocols.io/view/visual-frailty-in-the-ageing-population-a-scoping-dbra2m2e
Godfrey Wanok, Emilie McSwiggan, Vasilis Raptis, Baljean Dhillon, Ian Underwood, Peter Cackett
TITLE: Visual frailty in the ageing population: A scoping review AUTHORS: Godfrey Wanok, Emilie McSwiggan, Vasilis Raptis, Baljean Dhillon, Ian Underwood, Peter Cackett [DESCRIPTION] Visual frailty is a condition which has not been comprehensively explored by researchers and other professionals. This scoping review wi...
["[Review questions] What is the definition of visual frailty?\nWhat domains can be used in the assessment of the condition?\nAre there assessment tools for the condition in the ageing\npopulation?\nWhat factors are associated with visual frailty in the ageing population?\nWhat is the relationship between frailty and A...
50,160
Acupuncture for glucose and lipid metabolic disorders of polycystic ovarian syndrome: A systematic review protocol
1
dx.doi.org/10.17504/protocols.io.bu8qnzvw
https://www.protocols.io/view/acupuncture-for-glucose-and-lipid-metabolic-disord-bu8qnzvw
Yang Wu, Tao Peng, Yu Chen, Li Huang, Bisong He, Shaobin Wei
TITLE: Acupuncture for glucose and lipid metabolic disorders of polycystic ovarian syndrome: A systematic review protocol AUTHORS: Yang Wu, Tao Peng, Yu Chen, Li Huang, Bisong He, Shaobin Wei [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:justify"><a href=...
["Study registration and designThesystematic review protocol has been registered prospectively in the International Prospective Register of Systematic Reviews (PROSPERO) (CRD42020177846). This protocol is reported in line with the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols (PRISMA-P) st...
24,740
Luminex Bead Conjugation
null
dx.doi.org/10.17504/protocols.io.4ecgtaw
null
Andrew Crowley
TITLE: Luminex Bead Conjugation AUTHORS: Andrew Crowley [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for amine-coupling of proteins to magnetic Luminex beads</div><div class = "text-block"><span>Quantity as written: 1 x 10</span><span style = "vertical-align:super;">6</span><span> beads ...
["[Activation]\nResuspend beads by briefly vortexing; dispense\n[(contains approx. 1.25 million beads)]", "[Activation]\nBEAD SEPARATION BLOCK", "[Activation]\nCollect volume by pulse centrifugation", "[Activation]\nPlace on magnetic tube rack, shielded from light\nProtocol tipPartially open the microcentrifuge tube be...
47,359
Analysis of genetic relatedness and paternity assignment in wild Guinea baboons (Papio papio) based on microsatellites
1
dx.doi.org/10.17504/protocols.io.bsg7nbzn
https://www.protocols.io/view/analysis-of-genetic-relatedness-and-paternity-assi-bsg7nbzn
Federica Dal Pesco, Franziska Trede, Dietmar Zinner, Fischer Julia
TITLE: Analysis of genetic relatedness and paternity assignment in wild Guinea baboons (Papio papio) based on microsatellites AUTHORS: Federica Dal Pesco, Franziska Trede, Dietmar Zinner, Fischer Julia [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:justify...
["Sample collection and storage:\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\tWe collected samples for genetic analysis non-invasively conducting fecal sampling of all identified individuals...
86,461
Challenging Beam Test
1
dx.doi.org/10.17504/protocols.io.dm6gp33r5vzp/v1
https://www.protocols.io/view/challenging-beam-test-cyn5xvg6
Sabina Marciano, Roberta Marongiu
TITLE: Challenging Beam Test AUTHORS: Sabina Marciano, Roberta Marongiu [DESCRIPTION] The challenging beam test is used to measure motor performance, coordination, and balance [STEPS] SECTION: Setup 1. Lay a bench pad on the table and place the beam on top of the two empty cages ~3-4 cm of overhang. SECTION: Setup 2...
["[Setup] Lay a bench pad on the table and place the beam on top of the two empty cages ~3-4 cm of overhang.", "[Setup] Place the test subject’s home cage on the narrow end sideways so that the bream rests on top of the side wall.", "[Setup] Clean the beam with 70% ethanol and dry thoroughly before starting and after c...
25,703
08 Exploration of expression condition
null
dx.doi.org/10.17504/protocols.io.5cfg2tn
null
TJUSLS China
TITLE: 08 Exploration of expression condition AUTHORS: TJUSLS China [STEPS] ?. Transform the plasmid into bacteria used to express target protein(e.g. E.coli BL21(DE3)). ?. Take monoclone in the culture plate into LB tube and cultivate in shaking incubator overnight(10-12h) to activate bacteria. ?. Test the OD600 numb...
["Transform the plasmid into bacteria used to express target protein(e.g. E.coli BL21(DE3)).", "Take monoclone in the culture plate into LB tube and cultivate in shaking incubator overnight(10-12h) to activate bacteria.", "Test the OD600 number of bacteria, then pipe 5-10ul into each new 5 mL LB tube. Don’t forget to a...
40,293
How to prepare zebrafish brain tissue samples for biochemical assays
6
dx.doi.org/10.17504/protocols.io.bjkdkks6
https://www.protocols.io/view/how-to-prepare-zebrafish-brain-tissue-samples-for-bjkdkks6
Adrieli Sachett, Matheus Gallas-Lopes, Radharani , Greicy M M Conterato, Ana Herrmann, Angelo Piato
TITLE: How to prepare zebrafish brain tissue samples for biochemical assays AUTHORS: Adrieli Sachett, Matheus Gallas-Lopes, Radharani , Greicy M M Conterato, Ana Herrmann, Angelo Piato [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Zebrafish are increasingly used as a model animal in neuroscience r...
["[Preparations to collect animal tissue]\nBefore starting to collect animal tissue, it is important to prepare some settings in order to guarantee the appropriate preservation of the sample and, ultimately, the assessment of biochemical parameters;", "[Sample collection and processing]\nThe following steps should be c...
100,932
Senescent Cell Evaluations in Normal Tissues (SCENT) Mapping Center Bronchoscopy Protocol
0
null
https://www.protocols.io/view/senescent-cell-evaluations-in-normal-tissues-scent-detc3eiw
Dr. Patty j Lee, Dr. Monica Kraft, Dr. Andrew Nixon
TITLE: Senescent Cell Evaluations in Normal Tissues (SCENT) Mapping Center Bronchoscopy Protocol AUTHORS: Dr. Patty j Lee, Dr. Monica Kraft, Dr. Andrew Nixon [DESCRIPTION] Cellular senescence is a stress-response, as well as a critical component of cell fate during development, repair, resilience, and normal aging. D...
["[Goal and Objective] This project aims to collect, handle, store, and allocate normal healthy lung tissues and biofluids for constructing cellular senescence maps according to the standards established by the Steering Committee of the SenNet consortium. It seeks to identify senescent cell differences across the body,...
24,699
Agar Plate Preparation
null
dx.doi.org/10.17504/protocols.io.4c3gsyn
null
Andrew Crowley
TITLE: Agar Plate Preparation AUTHORS: Andrew Crowley [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Default scale - 1 L (makes approx. 2.5 sleeves of plates)</div></div> [STEPS] ?. Add powdered LB ( ) and agar () to deionized water ( ) 25 g 15 g 1 L n.b.Particulates may remain after addition, but...
["Add powdered LB ( ) and agar () to deionized water ( )\n25 g\n15 g\n1 L\nn.b.Particulates may remain after addition, but these will dissolve during the autoclave cycle", "Autoclave using liquid cycle", "Allow media to cool to\n50 °C", "Select antibiotic from the list below:", "Add kanamycin and mix thoroughly\n50 mg...
57,457
MicroStone: Exploring the capabilities of the Artec Micro in scanning stone tools
1
dx.doi.org/10.17504/protocols.io.81wgb6781lpk/v1
https://www.protocols.io/view/microstone-exploring-the-capabilities-of-the-artec-b4crqsv6
Armando Falcucci
TITLE: MicroStone: Exploring the capabilities of the Artec Micro in scanning stone tools AUTHORS: Armando Falcucci [DESCRIPTION] In this protocol, I describe how to scan small-sized lithics with the high-precision desktop 3D scanner Artec Micro. This scanner is particularly efficient when it comes to scan small objec...
["[Part 1 - Object positioning] Use the Vise Jig provided by Artec to securely hold the artifact between its jaws (Fig. 1). The Vise Jig can be safely mounted thanks to the magnetic plate (i.e., the Gum Jig) on the Screw Jig. The object should be stable enough to avoid movements during the scanning process. I recommend...
null
null
null
dx.doi.org/10.17504/protocols.io.exybfpw
null
null
TITLE: No Title AUTHORS: [STEPS] ?.
[]
80,277
RNA extraction from hairy roots of common bean (Phaseolus vulgaris L.) and cDNA synthesis
4
dx.doi.org/10.17504/protocols.io.8epv5jq24l1b/v1
https://www.protocols.io/view/rna-extraction-from-hairy-roots-of-common-bean-pha-csmvwc66
Ronal Pacheco Sánchez, Noreide Nava
TITLE: RNA extraction from hairy roots of common bean (Phaseolus vulgaris L.) and cDNA synthesis AUTHORS: Ronal Pacheco Sánchez, Noreide Nava [DESCRIPTION] Extracting RNA for subsequent quantification of transcript levels by RT-qPCR requires high purity and concentration. When the amount of tissue is not abundant, as ...
["[Extraction of total RNA] Macerate root tissue using liquid nitrogen.", "[Extraction of total RNA] Load 100 mg of macerated tissue into a 1.5 mL Eppendorf tube and add 1 mL of .", "[Extraction of total RNA] Mix by vortexing15 s and incubate for 5 min at room temperature.", "Centrifuge 11800 rpm, 15 min, 4 °C", "[...
20,730
Lipid Biomarker Extraction and Elution into different Fractions from sediment
null
dx.doi.org/10.17504/protocols.io.yg2ftye
null
Lucia Leierer, Antonio V. Herrera-Herrera, Margarita Jambrina-Enríquez
TITLE: Lipid Biomarker Extraction and Elution into different Fractions from sediment AUTHORS: Lucia Leierer, Antonio V. Herrera-Herrera, Margarita Jambrina-Enríquez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a protocol of a method used to obtain the total lipid extract (TLE) from archae...
["[Preparation]\nOven drying of sediment samples at during and subsampling to of homogenized sediment\n5 g", "[Lipid Extraction]\nAdd dichloromethane/methanol (DCM/MeOH) 9:1 v/v to the sediment\n40 ml", "[Lipid Extraction]\nMix for in a sonicator (below )", "[Lipid Extraction]\nTransfer the solvent into a centrifuge t...
102,123
Environmental DNA Metabarcoding- Full Pipeline Collection
0
dx.doi.org/10.17504/protocols.io.eq2lywxwpvx9/v1
https://www.protocols.io/view/environmental-dna-metabarcoding-full-pipeline-coll-dfyj3pun
Colleen Kellogg, Matt Lemay, rute.carvalho Carvalho, Andreas Novotny
TITLE: Environmental DNA Metabarcoding- Full Pipeline Collection AUTHORS: Colleen Kellogg, Matt Lemay, rute.carvalho Carvalho, Andreas Novotny [DESCRIPTION] This is a collection of protocols used to analyze environmental DNA (eDNA) of seawater samples. As part of the Hakai Institute Ocean Observing Program, biomolecul...
[]
65,182
TestoUltra Brief Information [REVIEWS]: SCAM &amp; LEGIT
1
dx.doi.org/10.17504/protocols.io.6qpvr6ydbvmk/v1
https://www.protocols.io/view/testoultra-brief-information-reviews-scam-amp-legi-cbv6sn9e
testolutraty
TITLE: TestoUltra Brief Information [REVIEWS]: SCAM &amp; LEGIT AUTHORS: testolutraty [DESCRIPTION] Getting more settled can cause a few troublesome issues with your sexual concurrence. That is the explanation we really want to edify you concerning another upgrade called TestoUltra Make Enhancement pills. One thing ...
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62,682
Small molecules released from islets of Langerhans determined by liquid chromatography – mass spectrometry
4
dx.doi.org/10.17504/protocols.io.6qpvr6rnzvmk/v1
https://www.protocols.io/view/small-molecules-released-from-islets-of-langerhans-b9f2r3qe
Emmanuel O. Ogunkunle, Matthew J. Donohue, Daniel J. Steyer, Damilola I. Adeoye, Wesley J. Eaton, Michael Roper
TITLE: Small molecules released from islets of Langerhans determined by liquid chromatography – mass spectrometry AUTHORS: Emmanuel O. Ogunkunle, Matthew J. Donohue, Daniel J. Steyer, Damilola I. Adeoye, Wesley J. Eaton, Michael Roper [DESCRIPTION] Islets of Langerhans are the endocrine tissue within the pancreas tha...
["[Methods] Benzoyl chloride derivatization\n\nFor production of calibration curves, 100 µLof a standard mixture of small molecules (concentrations given in Section 3.3 in the original publication) in BSS was mixed with 50 µL of 100 mM sodium carbonate (pH 9.2) and 50 µL 2% BzCl (by volume in ACN). The mixture was vort...
65,098
Ikaria Lean Belly Juice Customer Reviews and Complaints!
1
dx.doi.org/10.17504/protocols.io.rm7vzyem5lx1/v1
https://www.protocols.io/view/ikaria-lean-belly-juice-customer-reviews-and-compl-cbtisnke
komalyo
TITLE: Ikaria Lean Belly Juice Customer Reviews and Complaints! AUTHORS: komalyo [DESCRIPTION] Everybody has the right to have a decent body shape and feel happy with just being themselves. It is in every case great to see your appearance in the mirror and be happy with your actual appearance. Sadly, few out of ...
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64,680
In situ high-speed brightfield imaging for studies of aquatic organisms
4
null
https://www.protocols.io/view/in-situ-high-speed-brightfield-imaging-for-studies-cbegsjbw
Sean P. Colin, Brad J. Gemmell, John H. Costello, Kelly R R Sutherland
TITLE: In situ high-speed brightfield imaging for studies of aquatic organisms AUTHORS: Sean P. Colin, Brad J. Gemmell, John H. Costello, Kelly R R Sutherland [DESCRIPTION] Behavioral measurements of fragile aquatic organisms require specialized in situ techniques.We developed an in situ brightfield camera set-up ...
["[Select field site] This system is lightweight, compact and portable and can be used SCUBA diving from shore, docks or boats.The camera system can also potentially be towed vertically from a research vessel.", "[Assemble equipment] The brightfield camera system relies entirely on available off the shelf components an...
33,893
Assembling Algal Shaker
null
dx.doi.org/10.17504/protocols.io.bdcdi2s6
null
Jakub Nedbal
TITLE: Assembling Algal Shaker AUTHORS: Jakub Nedbal [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol described the final assembly steps to complete the illuminated orbital shaker and its testing. The steps involve fixing the transparent platform to the orbital shaker, placing the LED i...
["Prepare a tidy workbench with sufficient space and illumination to minimize risk of accidents.", "Fix the four M4×50 mm male-female standoffs to the orbital shaker platform using nuts and washers.Where Imperial threads are preferred, use #8-32×2\" stand-offs instead.​", "Lay the cooled LED illuminator onto the orbita...
62,438
Lanthanum DAB metals, Ln-DAB2 labeling of APEX2
1
dx.doi.org/10.17504/protocols.io.e6nvwkoe2vmk/v1
https://www.protocols.io/view/lanthanum-dab-metals-ln-dab2-labeling-of-apex2-b88erzte
Stephen R. Adams, Mason R. Mackey, Alice Ting, Mark Ellisman, Thomas Deerinck
TITLE: Lanthanum DAB metals, Ln-DAB2 labeling of APEX2 AUTHORS: Stephen R. Adams, Mason R. Mackey, Alice Ting, Mark Ellisman, Thomas Deerinck [DESCRIPTION] This protocol uses APEX2, an engineered peroxidase that genetically targets a cellular region of interest, to oxidatively polymerize Ln-DAB2, (lanthanum chelates c...
["HEK293T cells are cultured on imaging plates containing poly-d-lysine coated glass bottom No. 0 coverslips (P35GC-0-14C, MatTek Corporation).", "Cells are transiently transfected with either APEX2-H2B or mitochondrial matrix-APEX2 fusion using Lipofectamine 3000 (Life Technologies). APEX2 is fused to N-terminal of H2...
72,464
Electroporation Transformation Protocol in S. Cerevisiae
4
null
https://www.protocols.io/view/electroporation-transformation-protocol-in-s-cerev-cizquf5w
Stephanie Hood
TITLE: Electroporation Transformation Protocol in S. Cerevisiae AUTHORS: Stephanie Hood [DESCRIPTION] Protocol adapted by Stephanie Hood, 09/2022, from: An improved yeast transformation method for the generation of very large human antibody libraries Overview: This protocol calls for ~800-900 ul of cell pellet and...
["[2-3 Days before starting experiment] Streak out yeast strains for electroporation protocol.\nDepending on how many cultures you are setting up you may need to streak more than 1 plate to get enough isolated colonies. Grow at 30°C for 2 days.", "[Day before experiment] Grow 5 ml culture(s) of S. cerevisiae cells over...
61,452
Slant board seed gemination and transplanting for hydroponic research
4
null
https://www.protocols.io/view/slant-board-seed-gemination-and-transplanting-for-b79krr4w
noah.langenfeld , Bruce Bugbee
TITLE: Slant board seed gemination and transplanting for hydroponic research AUTHORS: noah.langenfeld , Bruce Bugbee [DESCRIPTION] Uniform seed germination is essential for hydroponic research. Conventional seed germination uses substrates such as peat moss or mineral wool. These substrates are difficult to remove f...
["[Seeding] Obtain a clean acrylic slant board and lay flat on a hard, clean surface.", "[Seeding] Cut a piece of germination paper 10 cm in height and the same length as the slant board.", "[Seeding] Place germination paper on top of slant board.", "[Seeding] Place seeds on top of germination paper (about 9 cm from th...
63,160
Deposition of matrix using an M5 TM sprayer for high resolution MALDI analysis
1
null
https://www.protocols.io/view/deposition-of-matrix-using-an-m5-tm-sprayer-for-hi-b9wyr7fw
Angela Kruse, Martin Dufresne, Jamie Allen, Danielle Gutierrez, Jeff Spraggins
TITLE: Deposition of matrix using an M5 TM sprayer for high resolution MALDI analysis AUTHORS: Angela Kruse, Martin Dufresne, Jamie Allen, Danielle Gutierrez, Jeff Spraggins [DESCRIPTION] This protocol describes the use of an HTX M5 TM sprayer to deposit small molecule matrices onto tissue sections for high reso...
["[Autofluorescence Scan] Remove slides from freezer and place in desiccator for 30 min", "[Autofluorescence Scan] Collect autofluorescence scan using a Zeiss AxioScan Z1 (https://www.protocols.io/edit/autofluorescence-microscopy-data-acquisition-b39kqr4w)", "[TM Sprayer Setup] Set nitrogen flow to 8 psi. turn on TM sp...
100,673
Sucrose lysis buffer
1
dx.doi.org/10.17504/protocols.io.j8nlk8op1l5r/v1
https://www.protocols.io/view/sucrose-lysis-buffer-dei93ch6
Colleen Kellogg
TITLE: Sucrose lysis buffer AUTHORS: Colleen Kellogg [DESCRIPTION] This protocol describes the preparation of sucrose lysis buffer to preserve DNA on sterivex filters. As part of the Hakai Institute Ocean Observing Program, biomolecular samples have been collected weekly, from 0 m to near bottom (260 m), to geneticall...
["[Methods] Filter-sterilize and label bottle.", "[Methods] Top up water to 500 mL. (no need to pH this one!)", "[Methods] Add a stir bar and dissolve all the powder.", "[Methods] Add milliQ water to about the 400 mL line.", "[Methods] Add 25mL of 1M Tris to the beaker.", "[Methods] Add 40 mL of 0.5M EDTA to the beaker...
25,511
Dispensing agar into multiwell plates July 2019 updates
null
dx.doi.org/10.17504/protocols.io.46fgzbn
null
Ida Barlow
TITLE: Dispensing agar into multiwell plates July 2019 updates AUTHORS: Ida Barlow [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Protocol for pouring agar into 96 well plates using Integra VIAFILL dispenser. Agar should be prepared in advance, and kept in 60</span><span style = "vertical-ali...
["[Configure Integra VIAFILL]\nInsert large cassette (suitable for dispensing volumes from 5-9999μL) into the machine:", "[Configure Integra VIAFILL]\nConfigure X, Y, and Z settings for the multiwell plate by clicking on tool symbol -> stage alignment -> 96 well 8Ch.\nFor UNIPLATE96SQWLF 650U:X = 95.6Y = 4.2Z = -22.5",...
49,485
Systemic AAV vectors for widespread and targeted gene delivery in rodents
3
dx.doi.org/10.17504/protocols.io.bujmnuk6
https://www.protocols.io/view/systemic-aav-vectors-for-widespread-and-targeted-g-bujmnuk6
Rosemary C. Challis, Sripriya Ravindra Kumar, Ken Y. Chan, Collin Challis, Keith Beadle, Min J. Jang, Hyun Min Kim, Pradeep S. Rajendran, John D. Tompkins, Kalyanam Shivkumar, Benjamin E. Deverman, Viviana Gradinaru
TITLE: Systemic AAV vectors for widespread and targeted gene delivery in rodents AUTHORS: Rosemary C. Challis, Sripriya Ravindra Kumar, Ken Y. Chan, Collin Challis, Keith Beadle, Min J. Jang, Hyun Min Kim, Pradeep S. Rajendran, John D. Tompkins, Kalyanam Shivkumar, Benjamin E. Deverman, Viviana Gradinaru [DESCRIPTION]...
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40,851
Background and Rationale (Part 1 of Phase 3 study of Vaccine Candidate for COVID-19)
1
dx.doi.org/10.17504/protocols.io.bj5tkq6n
https://www.protocols.io/view/background-and-rationale-part-1-of-phase-3-study-o-bj5tkq6n
Chris Ockenhouse, Chris Gast, Renee Holt, Jorge Flores
TITLE: Background and Rationale (Part 1 of Phase 3 study of Vaccine Candidate for COVID-19) AUTHORS: Chris Ockenhouse, Chris Gast, Renee Holt, Jorge Flores [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This is a collection of protocols for: "Phase 3 randomized, double-blinded, placebo-contro...
[]
102,395
Lysosomal Membrane Permeability (LMP) assay
0
dx.doi.org/10.17504/protocols.io.eq2lywxbevx9/v1
https://www.protocols.io/view/lysosomal-membrane-permeability-lmp-assay-df833ryn
Scott Vermilyea
TITLE: Lysosomal Membrane Permeability (LMP) assay AUTHORS: Scott Vermilyea [DESCRIPTION] This protocol details the requirements of the Lysosomal Membrane Permeability (LMP) assay. [STEPS] SECTION: Cell Culture/Treatments: 1. 0.5x10^4/ml of cells plated on coverslips. SECTION: Imaging: 6. Leica Stellaries 8 confocal ...
["[Cell Culture/Treatments:] 0.5x10^4/ml of cells plated on coverslips.", "[Imaging:] Leica Stellaries 8 confocal laser scanning microscope.", "[Cell Culture/Treatments:] PFF-488 treatment.", "[Cell Culture/Treatments:] PBS/trypsin (0.01%) wash.", "[Cell Culture/Treatments:] Fix cells using 4% PFA.", "[Cell Culture/Tre...
null
null
null
dx.doi.org/10.17504/protocols.io.jskcncw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
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48,412
Shipping Embryos in a Portable Incubator
4
dx.doi.org/10.17504/protocols.io.bth4nj8w
https://www.protocols.io/view/shipping-embryos-in-a-portable-incubator-bth4nj8w
Eliab Estrada Cortés, Peter Hansen
TITLE: Shipping Embryos in a Portable Incubator AUTHORS: Eliab Estrada Cortés, Peter Hansen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a protocol to describe how to ship non-frozen embryos</div></div> [STEPS] ?. [ ] Test the shipping incubator and make sure that it can remain powered f...
["[ ]\nTest the shipping incubator and make sure that it can remain powered for at least 24 hours while operating with the battery. We currently use TO 42 from WTA https://www.wtavet.com.br/?lang=en but others work well also. The temperature is set to 38.5°C", "[ ]\nStart charging shipping incubator 1 day before shippi...
67,586
SMRT-Tag - Sensitive multimodal profiling of native DNA by transposase-mediated single-molecule sequencing
4
dx.doi.org/10.17504/protocols.io.e6nvwk3b9vmk/v1
https://www.protocols.io/view/smrt-tag-sensitive-multimodal-profiling-of-native-cd9as92e
Scott Nanda, Ke Wu, Siva Kasinathan, vijay.ramani
TITLE: SMRT-Tag - Sensitive multimodal profiling of native DNA by transposase-mediated single-molecule sequencing AUTHORS: Scott Nanda, Ke Wu, Siva Kasinathan, vijay.ramani [DESCRIPTION] Here we describe a protocol for SMRT-Tag: Single-Molecule Real-Time sequencing by Tagmentation - a transposase-mediated strategy f...
["[gDNA QC] High Molecular Weight genomic DNA (HMW gDNA) is used as standard input for the SMRT-Tag method. \n\nTo reduce its viscosity, incubate at 37ºC for 5-10 min at low agitation speed (300 rpm). Pipette up and down 5–10 times using a p200 wide-bore pipette to ensure any clumps of DNA are dispersed.\n\n[OPTIONAL] ...
null
null
null
dx.doi.org/10.17504/protocols.io.rwcd7aw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol describing fluorescence immunohistochemistry for detection of thymosin beta 4 alongside CD31, smooth muscle actin and myosin heavy chain in PFA-fixed human cardiac tissue samples.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?.
["Tissue was fixed overnight in 4% PFA and then dehydrated the following day with 2 hour washes of 70%, 80%, 90% and 100% ethanol. Tissue was then placed in 100% chloroform overnight. The following day the tissue was embedded in paraffin wax, allowed to set and then stored at 4ºC.", "Using a Leica rotary microtome, 10 ...
72,184
3&#39;-Biotinylation of mRNA
1
null
https://www.protocols.io/view/3-39-biotinylation-of-mrna-ciqyudxw
Clark Fritsch
TITLE: 3&#39;-Biotinylation of mRNA AUTHORS: Clark Fritsch [DESCRIPTION] This protocol is meant to efficiently biotinylate the 3'-end of an mRNA prepared for use in single-molecule FRET experiments. The biotinylated 3'-end of the mRNA is able to bind tightly to streptavidin or neutravidin for use in the stopped-flow c...
["[Reagent Preparation] 1 mL 1M Na-acetate (pH=5.0):Add 52.5mg (MW 82.03) Na-acetate (64%) and 20.7 uL of 17.4 M Acetic acid (36%) in 800 uL of DEPC water, then make up the volume upto 1 ml by adding more DEPC water or you can use the purchased 3M Na-acetate (pH=5.2) and make it to 1M.", "[Reagent Preparation] 3M KCl:A...
86,661
Overcoming problematic growth phenotypes in organoids from patients with monogenic GI disease
4
dx.doi.org/10.17504/protocols.io.dm6gp33ypvzp/v1
https://www.protocols.io/view/overcoming-problematic-growth-phenotypes-in-organo-cyvdxw26
Katlynn Bugda Gwilt, Jay Thiagarajah
TITLE: Overcoming problematic growth phenotypes in organoids from patients with monogenic GI disease AUTHORS: Katlynn Bugda Gwilt, Jay Thiagarajah [DESCRIPTION] Patient-derived organoids provide a unique model system to explore disease causing mutations ex vivo. By use of organoids from duodenal or colonic biopsies of...
["[Thawing Frozen Lines] 300 x g Centrifuge at 300 x g for 5 minutes", "[Thawing Frozen Lines] Aspirate media leaving all of the residual matrigel from freezing behind", "[Thawing Frozen Lines] Add 200ul of fresh matrigel to the organoid/matrigel pellet", "[Thawing Frozen Lines] Gently resuspend using P1000 and 1000ul ...
null
null
null
dx.doi.org/10.17504/protocols.io.e4abgse
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <p><strong>INTRODUCTION </strong></p> <p>Protein assays are routinely used in many research fields to estimate proteins in a vast array of buffers and conditions. A major problem for researchers is to select a protein assay from the vast selection on the market that is compatible...
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null
null
null
dx.doi.org/10.17504/protocols.io.p3tdqnn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Here we describe the standardised protocol used by the <a href="http://www.caboscience.org" target="_blank" rel="noopener noreferrer">Canadian Airborne Biodiversity Observatory</a> (CABO) to measure leaf water content and specific leaf area, using the <a href="http://regent.q...
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45,063
Quick Protocol for Monarch® Plasmid Miniprep Kit (NEB #T1010)
1
dx.doi.org/10.17504/protocols.io.bp9fmr3n
https://www.protocols.io/view/quick-protocol-for-monarch-plasmid-miniprep-kit-ne-bp9fmr3n
New England Biolabs
TITLE: Quick Protocol for Monarch® Plasmid Miniprep Kit (NEB #T1010) AUTHORS: New England Biolabs [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This is the quick version of the Monarch® Plasmid DNA Miniprep Kit Protocol (NEB #T1010). For the full protocol, please click </span><a href="https:...
["Pellet 1–5 ml (not to exceed 15 OD units) bacterial culture by centrifugation for 30 seconds. Discard supernatant.\n1.5 ml of culture is sufficient for most applications. Ensure cultures are not overgrown (12-16 hours is ideal).", "Resuspend pellet in 200 μl Plasmid Resuspension Buffer (B1).\nVortex or pipet to ensur...