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75,773
FBS heat inactivation
4
null
https://www.protocols.io/view/fbs-heat-inactivation-cm85u9y6
Andreas Sagen
TITLE: FBS heat inactivation AUTHORS: Andreas Sagen [DESCRIPTION] The objective of heat inactivation is to destroy complement activity in the serum without affecting the growth-promoting characteristics of the product. Removal of complement activity from serum, such as fetal bovine serum, is not required for most cell...
["[Inactivation] Preparation", "[Inactivation] Aliquot FBS into 50 mL or 100 mL units in a sterile environment", "[Inactivation] Prepare a water bath", "[Inactivation] Clean and add distilled water to an appropriate hight", "[Inactivation] FBS inactivation", "[Inactivation] Set temperature to 58 °C", "[Inactivation] Pr...
85,681
Total lipid content analysis for coral samples
4
dx.doi.org/10.17504/protocols.io.81wgbx681lpk/v1
https://www.protocols.io/view/total-lipid-content-analysis-for-coral-samples-cxwrxpd6
Keyla Plichon, Paola Furla
TITLE: Total lipid content analysis for coral samples AUTHORS: Keyla Plichon, Paola Furla [DESCRIPTION] This protocol aims to extract and quantify total lipidic content from frozen coral samples. This method is adapted from the extraction protocol of Rowan et al., 2020 and the sulfo-phospho-vanillin protocol on 96-wel...
["[Lipid extraction] Remove subsamples from the freeze dryer.\nTurn off the freeze-dryer and quickly tighten the screw caps to the vial to avoid moisture entering the subsample. Keep samples at -80°C until analysis.", "[Lipid quantifcation sulfo-phospho-vanillin method] Prepare a stock of cholesterol solution and prepa...
78,939
Tetiaroa Yellow Crazy Ant Sampling: Sugar and PB Protocol - 1/24/2023
4
dx.doi.org/10.17504/protocols.io.n2bvj81rxgk5/v1
https://www.protocols.io/view/tetiaroa-yellow-crazy-ant-sampling-sugar-and-pb-pr-crb3v2qn
Juliet Capriola, Kalani Alcala
TITLE: Tetiaroa Yellow Crazy Ant Sampling: Sugar and PB Protocol - 1/24/2023 AUTHORS: Juliet Capriola, Kalani Alcala [DESCRIPTION] Walked along a series of transects with ant sugar water and peanut butter vials. The experiment aimed to discover the extent of the invasive yellow crazy Ant ,Anoplolepis gracilipes, on Te...
["[Preparation] Prepare 25 sugar plastic vials by soaking a napkin in sugar water and placing at the bottom of the vial", "[Preparation] Divide the atoll into a series of transects labeled A-Z. Then on each transect create points 1-X. The points should create a grid on the island.", "[Preparation] Prepare 12 PB plastic...
null
null
null
dx.doi.org/10.17504/protocols.io.htrb6m6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
92,442
A polarized cell system amenable to subcellular resolution imaging of influenza virus infection
1
dx.doi.org/10.17504/protocols.io.36wgq3pmklk5/v1
https://www.protocols.io/view/a-polarized-cell-system-amenable-to-subcellular-re-c6h2zb8e
Brault Jean-Baptiste, Thouvenot Catherine, Cannata Serio Magda, Paisant Sylvain, Fernandes Julien, Gény David, Danglot Lydia, Mallet Adeline, Naffakh Nadia
TITLE: A polarized cell system amenable to subcellular resolution imaging of influenza virus infection AUTHORS: Brault Jean-Baptiste, Thouvenot Catherine, Cannata Serio Magda, Paisant Sylvain, Fernandes Julien, Gény David, Danglot Lydia, Mallet Adeline, Naffakh Nadia [DESCRIPTION] This protocol details a polarized ce...
["[SEEDING AND POLARIZATION OF CACO-2/TC7 CELLS ON CYTODEX 3 BEADS] Prepare the repellent layer and seed the Caco-2/TC7 cells", "[SEEDING AND POLARIZATION OF CACO-2/TC7 CELLS ON CYTODEX 3 BEADS] Dissolve the Gelrite solution (0.8% Gelrite + 0.1% MgSO4 x7H2O w/v in PBS) in the microwave oven.", "[SEEDING AND POLARIZATIO...
82,131
L3 stage C. elegans dissociation for FACS isolation and RNA-seq analysis of intestine-specific cells
4
dx.doi.org/10.17504/protocols.io.14egn7zwmv5d/v2
https://www.protocols.io/view/l3-stage-c-elegans-dissociation-for-facs-isolation-cuftwtnn
Robert TP Williams, Erin Osborne Nishimura
TITLE: L3 stage C. elegans dissociation for FACS isolation and RNA-seq analysis of intestine-specific cells AUTHORS: Robert TP Williams, Erin Osborne Nishimura [DESCRIPTION] This protocol is for generating a single cell suspension suitable for isolation of intestine-specific cells through Fluorescence Activated Cell S...
["[Before beginning] Prepare reagents in advance\n\n\nL15-10 Buffer: Mix 500 ml Leibovitz's L-15 Medium, 50 ml Fetal Bovine Serum (heat inactivated), 50 ul of 100x Penicillin-Streptomycin solution and 7.7 g sucrose. Filter with 0.2 micron pore filter. Store at 4ºC.\n\nEgg Buffer: Mix 29.5 ml of 2M NaCl, 12 ml of 2M KCl...
71,205
RNA-interference in Paramecium bursaria
2
dx.doi.org/10.17504/protocols.io.8epv5jzm4l1b/v1
https://www.protocols.io/view/rna-interference-in-paramecium-bursaria-chsdt6a6
Ben Jenkins
TITLE: RNA-interference in Paramecium bursaria AUTHORS: Ben Jenkins [DESCRIPTION] RNA-interference based gene knock-down in Paramecium bursaria. This collection contains protocols for: 1) Paramecium cultures for RNAi - Filtering and preparing P. bursaria cultures for downstream RNAi 2) RNAi induction - Back-dilution,...
[]
90,500
stilPCR (single-tube Illumina long read PCR) increases the effective sequencing length of Illumina, targeted next-generation sequencing
4
dx.doi.org/10.17504/protocols.io.bp2l6xmn5lqe/v1
https://www.protocols.io/view/stilpcr-single-tube-illumina-long-read-pcr-increas-c4mcyu2w
Jason D Limberis, Roland Nagel, Soumitesh Chakravorty, Dena Block, Scott Dewell, Alina Nalyvayko, Zach Howard, Grant Theron, Rouxjeane Venter, John Metcalfe
TITLE: stilPCR (single-tube Illumina long read PCR) increases the effective sequencing length of Illumina, targeted next-generation sequencing AUTHORS: Jason D Limberis, Roland Nagel, Soumitesh Chakravorty, Dena Block, Scott Dewell, Alina Nalyvayko, Zach Howard, Grant Theron, Rouxjeane Venter, John Metcalfe [DESCRIPTI...
["[Stage 1 PCR] COMPONENT FINAL C. 50ul 5X Reaction Buffer 1X 10 5X GC Buffer 1X 10 10 mM dNTPs 200 µM 1 AtpE_stilPCRm1_F1 0.1µM 0.5 AtpE_stilPCRm1_R1 0.1µM 0.5 Rv0678_stilPCRm1_F1 0.1µM 0.5 Rv0678_stilPCRm1_R2 0.1µM 0.5 Template DNA – 2 Q5 High-Fidelity DNA Polymerase 0....
98,739
A protocol for tissue clearing and three-dimensional imaging of human sigmoid mucosal biopsies
0
dx.doi.org/10.17504/protocols.io.6qpvr8dkblmk/v2
https://www.protocols.io/view/a-protocol-for-tissue-clearing-and-three-dimension-dcnt2ven
Pu-Qing Yuan, Tao Li, Yvette Taché
TITLE: A protocol for tissue clearing and three-dimensional imaging of human sigmoid mucosal biopsies AUTHORS: Pu-Qing Yuan, Tao Li, Yvette Taché [DESCRIPTION] This protocol was developed for tissue clearing and 3D imaging of human sigmoid mucosal biopsies by adapting and modifying the original CLARITY tissue clearin...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ssyeefw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol is used to clarity the process of total DNA extration for L. maculatus genome.</p> [STEPS] ?. ?. ?. ?. ?. ?.
[]
82,136
Implant Surgery: Chronic recoverable Neuropixels in mice
1
dx.doi.org/10.17504/protocols.io.yxmvmnn2bg3p/v6
https://www.protocols.click/view/implant-surgery-chronic-recoverable-neuropixels-in-cufywtpw
Emily A Aery Jones
TITLE: Implant Surgery: Chronic recoverable Neuropixels in mice AUTHORS: Emily A Aery Jones [DESCRIPTION] This protocol collection explains how to build a low-cost, lightweight system to implant 1 Neuropixels 1.0 probe or 2 Neuropixels 2.0 probes into mice, record during freely moving behavior, then recover the probes...
["[Prepare surgical tools and field] Sterilize tips of metal instruments, ground screw, and headbar in autoclave 25 min or hot bead sterilizer 5 s . Disinfect cotton swabs, toothpick, and a weigh boat under UV light 30 min .", "[Prepare surgical tools and field] Disinfect surgical field with 10% bleach. Lay absorbent ...
19,445
Adapted protocol to extract total RNA using TRIzol® (Invitrogen, Thermo)
null
dx.doi.org/10.17504/protocols.io.w8vfhw6
null
Talita Gagliardi
TITLE: Adapted protocol to extract total RNA using TRIzol® (Invitrogen, Thermo) AUTHORS: Talita Gagliardi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Total RNA extraction using TRizol reagent.</div><div class = "text-block">Attention: for nasopharyngeal aspirates, we use regular TRizol reagent i...
["Incubate the homogeneized sample for 5min at r.t.", "Add 150ul of chloroform", "Shake the tube vigorously by vortex for 15sec.", "Incubate for 3min at r.t.", "Centrifuge for 15min at 12,000 x g and 4ºC.", "The mixture separates into red lower phenol-chloroform phase, and interphase, and a colorless upper aqueous phas...
null
null
null
dx.doi.org/10.17504/protocols.io.esxbefn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] For use in <a href="https://www.protocols.io/view/CviRI-Purification-From-XZ-6E-Virus-Infected-NC64A-er4bd8w" target="_blank">CviRI Purification From XZ-6E Virus Infected NC64A Chlorella</a>. [STEPS] ?. ?. ?.
[]
27,578
Isolation of Klebsiella strains from human or animal fecal material
null
dx.doi.org/10.17504/protocols.io.662hhge
null
Sylvain Brisse, virginie passet, Carla Rodrigues
TITLE: Isolation of Klebsiella strains from human or animal fecal material AUTHORS: Sylvain Brisse, virginie passet, Carla Rodrigues [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is intended for isolation of Klebsiella strains from human or animal fecal material. It is derived from ...
["Pre-treatment of sampleEnrich the sample by use of an enrichment broth. We typically use Luria-Bertani broth with 10 mg/L of ampicillin final concentration. Because they carry a constitutively expressed class A beta-lactamase (blaSHV type), the vast majority of Klebsiella isolates will grow/survive, whereas many othe...
54,878
FPCount protocol - Short protocol
4
dx.doi.org/10.17504/protocols.io.bzt6p6re
https://www.protocols.io/view/fpcount-protocol-short-protocol-bzt6p6re
Eszter Csibra, Guy-Bart Stan
TITLE: FPCount protocol - Short protocol AUTHORS: Eszter Csibra, Guy-Bart Stan [DESCRIPTION] FPCount is a complete protocol for fluorescent protein calibration, consisting of: 1. FP expression/purification using Thermo's HisPur Cobalt Resin. 2. FP concentration determination in a microplate reader. 3. FP fluores...
["[Harvesting and Washing] [ Day 2 ]\n\nBuffers:\nWash buffer = T50N150\n 50 mM Tris-Cl pH 7.5, 150 mM NaCl\n Doesn’t need protease inhibitors\n Can be substituted by T50N300\nResuspension buffer = T50N300+pi\n 50 mM Tris-Cl pH 7.5, 300 mM NaCl\n protease inhibitors (pi; 1 tablet/10ml) \n filter steri...
44,314
Immunoprecipitation Washes and RNA Dephosphorylation
4
dx.doi.org/10.17504/protocols.io.bph2mj8e
https://www.protocols.io/view/immunoprecipitation-washes-and-rna-dephosphorylati-bph2mj8e
Eric L. Van Nostrand, Thai B. Nguyen, Chelsea Gelboin-Burkhart, Ruth Wang, Steven M. Blue, Gabriel A. Pratt, Ashley L. Louie, Gene W. Yeo
TITLE: Immunoprecipitation Washes and RNA Dephosphorylation AUTHORS: Eric L. Van Nostrand, Thai B. Nguyen, Chelsea Gelboin-Burkhart, Ruth Wang, Steven M. Blue, Gabriel A. Pratt, Ashley L. Louie, Gene W. Yeo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Profiling of RNA binding protein targets in v...
["[Wash Beads (Pre-chill All Wash Buffers to 4 °C)]\nMagnetically separate beads, remove the supernatant (or store for western if desired).\nWe often perform intermediate washes with equal mixtures of the previous and current wash buffer, in order to decrease dramatic changes in buffer composition.", "[Wash Beads (Pre-...
76,671
Isolation of mouse embryonic fibroblasts (MEFs) from mouse embryos at E12.5
4
dx.doi.org/10.17504/protocols.io.eq2ly713qlx9/v1
https://www.protocols.io/view/isolation-of-mouse-embryonic-fibroblasts-mefs-from-cn47vgzn
Francesca Tonelli
TITLE: Isolation of mouse embryonic fibroblasts (MEFs) from mouse embryos at E12.5 AUTHORS: Francesca Tonelli [DESCRIPTION] Mouse embryo fibroblasts (MEFs) isolated from genetically altered mouse strains are a powerful resource for investigating the consequences of defined genetic mutations at the cellular level. Here...
["Transfer the uterus containing the embryos to a dish with ice cold PBS.", "Separate the embryos by cutting through the uterus in the region between each embryo with scissors.", "Transfer each embryo to a new dish with ice cold PBS.", "Remove the placenta and other maternal tissues by gently pulling them off using for...
20,048
Transfection of primary endothelial cells with siRNA
null
dx.doi.org/10.17504/protocols.io.xtqfnmw
null
Thomas Mudersbach, Daniel Siuda, Karin Kohlstedt, Ingrid Fleming
TITLE: Transfection of primary endothelial cells with siRNA AUTHORS: Thomas Mudersbach, Daniel Siuda, Karin Kohlstedt, Ingrid Fleming [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Preparing the endothelial cells for transfection (12-well plate)] Cells were seeded the day before transfection at a density...
["[Preparing the endothelial cells for transfection (12-well plate)]\nCells were seeded the day before transfection at a density of 3 x 104 cells/cm2.", "[Transfection ]\nCells were washed twice with PBS and than starved for at least in serum-free basal MCDB131 Medium at before incubation is finshed prepare the transfe...
53,684
Homology modeling using Phyre2 for Biochemistry I
1
null
https://www.protocols.io/view/homology-modeling-using-phyre2-for-biochemistry-i-bynupvew
Michael Friedman, Chris Berndsen
TITLE: Homology modeling using Phyre2 for Biochemistry I AUTHORS: Michael Friedman, Chris Berndsen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for homology modeling proteins for use in Biochemistry I at James Madison University. Protocol guides students to use the PHYRE2 web server (cit...
["[NCBI BLAST]\nNavigate to NCBI BLAST (Basic Local Sequence Alignment Tool) and paste your sequence into the \"Enter Query Sequence\" box.", "[NCBI BLAST]\nThe standard settings for the search are shown in the table. ABC1Default SettingWhat it does2Enter Query Sequence3Query Subrange(Blank)Limits search to a part of ...
94,565
Purification of GST-LC3B
1
dx.doi.org/10.17504/protocols.io.3byl4qnbjvo5/v1
https://www.protocols.io/view/purification-of-gst-lc3b-c8kdzus6
Elias Adriaenssens
TITLE: Purification of GST-LC3B AUTHORS: Elias Adriaenssens [DESCRIPTION] This protocol describes the purification of GST-LC3B. [STEPS] SECTION: Purification of GST-LC3B 1. To purify GST-LC3B, insert human LC3B cDNA in a pGEX-4T1 vector. Note, the last five amino acids of LC3B are deleted to mimic the cleavage by ATG...
["[Purification of GST-LC3B] To purify GST-LC3B, insert human LC3B cDNA in a pGEX-4T1 vector. Note, the last five amino acids of LC3B are deleted to mimic the cleavage by ATG4.", "[Purification of GST-LC3B] After the transformation of the pGEX-4T1 vector encoding GST-LC3B in E.coli Rosetta (DE3) pLysS cells, grow the c...
null
null
null
dx.doi.org/10.17504/protocols.io.dm448v
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
26,075
Bradford protein assay – Protein concentration measurement (single 595 nm read)
1
dx.doi.org/10.17504/protocols.io.14egnxd3ml5d/v1
https://www.protocols.io/view/bradford-protein-assay-protein-concentration-measu-5p3g5qn
Daniel C Moreira
TITLE: Bradford protein assay – Protein concentration measurement (single 595 nm read) AUTHORS: Daniel C Moreira [DESCRIPTION] This protocol describes how to measure the concentration of total protein in a sample performing the Bradford's assay using microtiter plates. Procedures are slightly modifications based on t...
["[Bradford&#x27;s protein reagent preparation] Prepare a solution containing 0.01% (w/v) Coomassie Brilliant Blue G-250 (e.g., B0770, Sigma-Aldrich), 4.7% (v/v) ethanol, and 8.5% (v/v) phosphoric acid as described in the next steps.", "[Bradford&#x27;s protein reagent preparation] Weight 100 mg of Coomassie Brilliant ...
57,891
YAP1 mKate:BSD HDR Knock-in (via Cas9 RNP lipofection)
4
null
https://www.protocols.io/view/yap1-mkate-bsd-hdr-knock-in-via-cas9-rnp-lipofecti-b4sbqwan
Emir Bora Akmeriç
TITLE: YAP1 mKate:BSD HDR Knock-in (via Cas9 RNP lipofection) AUTHORS: Emir Bora Akmeriç [DESCRIPTION] s [STEPS] SECTION: Culturing Immortalized HUVECs 1. In a t25 flask, add 10 mL of EGM2 (with antibiotics). Incubate flask for 20 minutes SECTION: Culturing Immortalized HUVECs 1.1. Thaw a 500k/1M CI-huVEC tube in a...
["[Culturing Immortalized HUVECs] In a t25 flask, add 10 mL of EGM2 (with antibiotics). Incubate flask for 20 minutes", "[Culturing Immortalized HUVECs] Thaw a 500k/1M CI-huVEC tube in a 37C water bath\nand add the cells into the flask.", "[Culturing Immortalized HUVECs] Change media 6-8 hours later or when cells have ...
62,617
Prima United Kingdom | Boosts Level Of Power &amp; Increases Assurance!
3
dx.doi.org/10.17504/protocols.io.3byl4bx82vo5/v1
https://www.protocols.io/view/prima-united-kingdom-boosts-level-of-power-amp-inc-b9dzr276
prima
TITLE: Prima United Kingdom | Boosts Level Of Power &amp; Increases Assurance! AUTHORS: prima [DESCRIPTION] Prima United Kingdom [STEPS]
[]
90,668
Regional Mouse Brain Analysis (Modified QUINT)
2
dx.doi.org/10.17504/protocols.io.kqdg3xbkzg25/v1
https://www.protocols.io/view/regional-mouse-brain-analysis-modified-quint-c4skywcw
Michael X. Henderson
TITLE: Regional Mouse Brain Analysis (Modified QUINT) AUTHORS: Michael X. Henderson [DESCRIPTION] This is series of protocols that has been adapted from published and unpublished protocols broadly referred to as the QUINT workflow. Note that the original QUINT workflow was generated by Yates and colleagues, and all cr...
[]
11,431
Liposome binding assay
null
dx.doi.org/10.17504/protocols.io.pefdjbn
null
Christa Testerink, Anne Froyd-Rankenberg
TITLE: Liposome binding assay AUTHORS: Christa Testerink, Anne Froyd-Rankenberg [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Standard protocol to examine lipid binding of the proteins isolated from the E.coli with the GST tag</div></div> [STEPS] ?. Mix lipids in chloroform in the right molecular...
["Mix lipids in chloroform in the right molecular ratios. Per sample 400 nmole (total) lipid is required. For (poly) phosphoinositides add MeOH (C:M 20:9)", "Dry lipid mixtures for 30’ in the speed vac. Store at -20C", "Add 500ul extrusion buffer per tube with lipid mixture. Vortex and let lipids rehydrate for 1-1.5 h ...
62,294
Superior Nutra Keto
3
dx.doi.org/10.17504/protocols.io.x54v9ykjzg3e/v1
https://www.protocols.io/view/superior-nutra-keto-b83wrype
H H
TITLE: Superior Nutra Keto AUTHORS: H H [DESCRIPTION] Superior Nutra Keto are most commonly offered in conjunction with a high-deductible health plan. As a rule, these plans keep premium costs low [STEPS]
[]
75,228
Preparing MEF-cultured hPSCs for nucleofection
4
dx.doi.org/10.17504/protocols.io.bp2l611ddvqe/v2
https://www.protocols.io/view/preparing-mef-cultured-hpscs-for-nucleofection-cmp4u5qw
Hanqin Li, Oriol Busquets, Steven Poser, Dirk Hockemeyer, Frank Soldner
TITLE: Preparing MEF-cultured hPSCs for nucleofection AUTHORS: Hanqin Li, Oriol Busquets, Steven Poser, Dirk Hockemeyer, Frank Soldner [DESCRIPTION] This protocol describes the procedure or preparing MEF-cultured human pluripotent stem cells (hPSCs) for the delivery of plasmids, mRNA or ribonucleoprotein (RNP) using ...
["When MEF-cultured hPSCs reach 50% confluency, change medium to hPSCs medium + Rock inhibitor, preparing for nucleofection the next day. For each 20 µl nucleofection reaction, prepare half to 1 well of cells.\n\nA detailed protocol on maintaining MEF-cultured hPSCs can be found in the collection \"Maintenance and inac...
20,493
Western blot analysis
null
dx.doi.org/10.17504/protocols.io.x9mfr46
null
Kiichi Hirota
TITLE: Western blot analysis AUTHORS: Kiichi Hirota [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Cell lysis] Scrape cells and transfer the suspension to a 1.5 ml microcentrifuge tube. ?. [Cell lysis] Centrifuge at 10,000 x g for 5 minutes at 4°C and collect the supernatant in a new microcentrifuge tube...
["[Cell lysis]\nScrape cells and transfer the suspension to a 1.5 ml microcentrifuge tube.", "[Cell lysis]\nCentrifuge at 10,000 x g for 5 minutes at 4°C and collect the supernatant in a new microcentrifuge tube.", "[Cell lysis]\nDetermine protein concentration by using DC™ Protein Assay Kit.", "[Separation of proteins...
null
null
null
dx.doi.org/10.17504/protocols.io.rhnd35e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>End-stage renal disease (ESRD) is related to high morbidity, mortality, and impaired health-related quality of life. While hemodialysis (HD) is the current life-saving standard of treatment for patients with ESRD, their quality of life (QoL) remains far from desirable. Online...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.pumdnu6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol describes how to make chemically competent <em>Vibrio natriegens</em> cells.</p> <p>The protocol was described and published by Weinstock et al., 2016</p> [BEFORE_START] <p>Make sure you have all solutions there:</p> <ul> <li><strong>Brain heart infusion + V<su...
[]
35,786
ddPCR Titration of Lentivirus Vectors
1
dx.doi.org/10.17504/protocols.io.be7ijhke
https://www.protocols.io/view/ddpcr-titration-of-lentivirus-vectors-be7ijhke
Addgene The Nonprofit Plasmid Repository
TITLE: ddPCR Titration of Lentivirus Vectors AUTHORS: Addgene The Nonprofit Plasmid Repository [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol describes ddPCR titration of Lentivirus vectors. To see the full abstract and additional resources, visit </span><a href="https://www.add...
["[Transducing Cells]\nThaw the virus.\non ice", "[Transducing Cells]\nPrepare the following dilution series of virus in DMEM complete: ABCD1Dilution FactorVirusMediaTotal Volume22.5160µL viral stock240µL400µL35200µL of 2.5-fold dilution200µL400µL410200µL of 5-fold dilution 200µL400µL520200µL of 10-fold dilution200µL...
104,832
Mouse brain, gut and plasma collection
0
dx.doi.org/10.17504/protocols.io.14egn3pkzl5d/v2
https://www.protocols.io/view/mouse-brain-gut-and-plasma-collection-dik84czw
Livia Hecke Morais
TITLE: Mouse brain, gut and plasma collection AUTHORS: Livia Hecke Morais [DESCRIPTION] Protocol used in the Mazmanian lab for collecting brain and gut tissues and plasma from mouse for metabolomics. Protocol has been approved by the California Institute of Technology’s Institutional Animal Care and Use Committee (IA...
["[Mouse brain, gut and plasma collection] At 4 months of age, mice is euthanized inside chemical hood by decapitation using scissors (as approved by IACUC protocol)", "[Mouse brain, gut and plasma collection] The head is separated from the rest of the body for brain and gut dissections using straight scissors", "[Mous...
null
null
null
dx.doi.org/10.17504/protocols.io.p4zdqx6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>We used the NEPA electroportor to transform Emiliania huxleyi cells. </p> <p>We were able to establish that this method can be used to introduce proteins into the cells. </p> [STEPS] ?. ?. ?. ?. ?. ?.
[]
23,772
Allende HRTEM study
null
dx.doi.org/10.17504/protocols.io.3f4gjqw
null
H. I. Cruz-Rosas, F. Riquelme, P. Santiago, L. Rendón, T. Buhse, F. Ortega-Gutiérrez, R. Borja-Urby, D. Mendoza, C. Gaona, P. Miramontes, G. Cocho.
TITLE: Allende HRTEM study AUTHORS: H. I. Cruz-Rosas, F. Riquelme, P. Santiago, L. Rendón, T. Buhse, F. Ortega-Gutiérrez, R. Borja-Urby, D. Mendoza, C. Gaona, P. Miramontes, G. Cocho. [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;"> </span><span style = "font-weight...
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69,180
Nasal Lavage Sample collection and processing
1
null
https://www.protocols.io/view/nasal-lavage-sample-collection-and-processing-cfs4tngw
maria.xylaki
TITLE: Nasal Lavage Sample collection and processing AUTHORS: maria.xylaki [DESCRIPTION] This protocol describes the nasal lavage fluid sampling that can be used for Omics (for Proteome analysis, see Schoenebeck et al, 2015; for Microbiome analysis see Heintz-Buschart et al, 2017) or RT-QuIC. Collection proceedure wa...
["[Nasal Lavage Sample collection] Two nasal lavage samples (NLS) are collected per person in each step, one from each nostril.\nBoth nasal cavities are pre-washed with 2 mL of physiological NaCl solution with the use of a plastic disposable pipette to remove bacterial impurities.", "[Nasal Lavage Sample collection] Af...
null
null
null
dx.doi.org/10.17504/protocols.io.qnmdvc6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Protocol for add azide modified nucleotide to 3' end of unmodified DNA olgionucleotide using terminal transferase. Protocol adapted from <a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4787804/" target="_blank" rel="noopener noreferrer">Winz et al. 2015</a>. </p> [STEP...
[]
48,928
Ethanol extraction protocol
1
null
https://www.protocols.io/view/ethanol-extraction-protocol-btz8np9w
Nannie
TITLE: Ethanol extraction protocol AUTHORS: Nannie [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Developed for IBA at the NRM DNA lab.</div></div> [STEPS] ?. Decant the ethanol through a mesh and add new ethanol to the insects. ?. Filter the ethanol: method 2.1 or 2.2 ?. Vacuum pumpPut the Steri...
["Decant the ethanol through a mesh and add new ethanol to the insects.", "Filter the ethanol: method 2.1 or 2.2", "Vacuum pumpPut the Sterivex filters on the vacuum pump with the large syringe attached to the filter. Turn on the pump and pour the ethanol into the syringe.The filtered ethanol will not be saved.", "Manu...
49,605
Cardiodynamic variables measured by Impedance Cardiography during a 6-minute walk test are reliable predictors of peak oxygen consumption in young healthy adults
1
dx.doi.org/10.17504/protocols.io.bupdnvi6
https://www.protocols.io/view/cardiodynamic-variables-measured-by-impedance-card-bupdnvi6
Fang Liu, Raymond Tsang, Alice YM Jones, Minchao Zhou, Kaiwen Xue, Miaoling Chen, Yulong Wang
TITLE: Cardiodynamic variables measured by Impedance Cardiography during a 6-minute walk test are reliable predictors of peak oxygen consumption in young healthy adults AUTHORS: Fang Liu, Raymond Tsang, Alice YM Jones, Minchao Zhou, Kaiwen Xue, Miaoling Chen, Yulong Wang [DESCRIPTION] <div class = "text-blocks"><div...
["Invitation emails were sent via the student portal of all students registered at the School of Rehabilitation of a local university affiliated with the hospital where this project was conducted.", "Exclusion criteria were: (1) students with known cardiovascular, pulmonary or musculoskeletal disorders that might limit...
29,169
ChroPlate - ProteinA/G
null
dx.doi.org/10.17504/protocols.io.8qrhvv6
null
David Frommholz, Alexandra Ehl, Nadine Stefanczyk
TITLE: ChroPlate - ProteinA/G AUTHORS: David Frommholz, Alexandra Ehl, Nadine Stefanczyk [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Purification Guide for the Isolation of Antibodies with ChroPlate Filtration Plates by DALEX Biotech.</span></div><div class = "t...
["[Load and Wash]\nPlace the ChroPlate on a clean deep-well plate. Add up to 1.5 ml sample to every well. Centrifuge 2 - 10 minutes at 400 g in a swing-out rotor.\nIt is advisable to keep the flow-through and wash fractions for later analysis, e.g. SDS-PAGE.The centrifugation time depends on the sample's volume and vis...
42,869
MRM-LC-MS/MS Assay for inflammatrory associated proteins in serum
1
dx.doi.org/10.17504/protocols.io.bm4vk8w6
https://www.protocols.io/view/mrm-lc-ms-ms-assay-for-inflammatrory-associated-pr-bm4vk8w6
TITLE: MRM-LC-MS/MS Assay for inflammatrory associated proteins in serum AUTHORS: [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Multiplex targeted proteomic assay to measure human proteins associated with inflammation in serum. A unique peptide is selected to represent a protein and is measured ...
["[Sample preparation]\nsamples are prepared for targeted mass spectrometric analysis utilising 20 μL serum.", "[Sample preparation]\nPrior to preparation, 5 pmol yeast enolase 1 protein (ENO1) (sigma UK) were added to each 20 μL serum sample to act as an internal standard, accounting for variation in sample preparati...
39,736
Materials and Methods: First report of the bat fly species Basilia italica in Romania
3
dx.doi.org/10.17504/protocols.io.bi2ykgfw
https://www.protocols.io/view/materials-and-methods-first-report-of-the-bat-fly-bi2ykgfw
aronpeter92 , Sándor D. Attila, Andrei D. Mihalca
TITLE: Materials and Methods: First report of the bat fly species Basilia italica in Romania AUTHORS: aronpeter92 , Sándor D. Attila, Andrei D. Mihalca [STEPS]
[]
71,801
Tissue Sectioning - Stanford
1
dx.doi.org/10.17504/protocols.io.j8nlkwkr1l5r/v1
https://www.protocols.io/view/tissue-sectioning-stanford-ciczuax6
emonte
TITLE: Tissue Sectioning - Stanford AUTHORS: emonte [DESCRIPTION] Tissue sectioning preparation (FF and FFPE) [STEPS] 1. Tissue sectioning protocol can be found here:
["Tissue sectioning protocol can be found here:"]
53,420
p2 21/09
1
null
https://www.protocols.io/view/p2-21-09-byekptcw
name go test surname
TITLE: p2 21/09 AUTHORS: name go test surname [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">test</div></div> [STEPS] ?. est
["est"]
53,638
Auger/Auspice/UShER SARS-CoV-2 Cluster Detection Workflow for the Terra Platform
5
null
https://www.protocols.io/view/auger-auspice-usher-sars-cov-2-cluster-detection-w-bymepu3e
Anusha Ginni, Jill V Hagey, Michael Weigand, Technical Outreach and Assistance for States Team
TITLE: Auger/Auspice/UShER SARS-CoV-2 Cluster Detection Workflow for the Terra Platform AUTHORS: Anusha Ginni, Jill V Hagey, Michael Weigand, Technical Outreach and Assistance for States Team [DESCRIPTION] The Titan_Augur workflow processes concatenated assembly fasta data for SARS-CoV-2 phylogenetic analysis and ...
["[Setup Terra and Google Cloud Accounts]", "[Import Augur workflows from Dockstore] Importing the Titan_Augur Workflows from Dockstore to the Terra User Workspace\n\nNote: The Augur workflow includes two steps: Titan_Augur_Prep and Titan_Augur_Run.\n\nThe Titan_Augur_Prep workflow prepares the metadata and assembly fa...
46,664
Laser Capture Microdissection 
4
null
https://www.protocols.io/view/laser-capture-microdissection-brtgm6jw
Angela Kruse, Carrie Romer, Jamie Allen, Danielle Gutierrez, Jeff Spraggins
TITLE: Laser Capture Microdissection  AUTHORS: Angela Kruse, Carrie Romer, Jamie Allen, Danielle Gutierrez, Jeff Spraggins [DESCRIPTION] Laser Capture Microdissection (LCM) is used to excise specific tissue regions for in-solution analysis. Tissue sections are mounted on membrane-coated slides, and a combination...
["[Turning on the instrument and software] Turn on the computer and microscope in the following order.", "[Loading a Sample] Load a sample mounted on a suitable membrane-coated slide.", "[Turning on the instrument and software] Turn key to ON position.", "[Turning on the instrument and software] Turn on computer.", "[T...
63,755
Mitochondrial Bioenergetics
4
dx.doi.org/10.17504/protocols.io.q26g746n3gwz/v1
https://www.protocols.io/view/mitochondrial-bioenergetics-cahjsb4n
mitsjoecohen
TITLE: Mitochondrial Bioenergetics AUTHORS: mitsjoecohen [DESCRIPTION] Mitochondrial bioenergetics plays a key role in a variety of basic cellular processes, such as energy production, nucleotide biosynthesis, and metabolism. Not only the basic system of cell production and death (apoptosis), it is also necessary fo...
[]
99,539
USDA LTAR Common Experiment measurement: Obtaining quality metrics in forage aboveground biomass
0
dx.doi.org/10.17504/protocols.io.n2bvjn6wpgk5/v1
https://www.protocols.io/view/usda-ltar-common-experiment-measurement-obtaining-ddft23nn
Kimberly A. Cassida, Harold Collins
TITLE: USDA LTAR Common Experiment measurement: Obtaining quality metrics in forage aboveground biomass AUTHORS: Kimberly A. Cassida, Harold Collins [DESCRIPTION] Harvesting crops for potential use as livestock forage must include nutritional value measurements to understand how these feedstuffs will meet the consumpt...
["[Sample collection] Sampling for forage quality requires the collection of biomass samples in addition to those samples outlined in the LTAR aboveground biomass protocol. These additional samples are necessary to capture a sample that reflects the cutting height of forage harvesting equipment or animal grazing. To be...
31,278
HuBMAP: Embedding Fixed Frozen OCT Samples
null
dx.doi.org/10.17504/protocols.io.basniede
null
Marda Jorgensen, Jerelyn Nick
TITLE: HuBMAP: Embedding Fixed Frozen OCT Samples AUTHORS: Marda Jorgensen, Jerelyn Nick [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The purpose of this Standard Operating Procedure (SOP) is to outline procedures for the OCT embedding of HuBMAP frozen fixed specimens.</div></div> [STEPS] ?. [Re...
["[Reagent Preparation]\nPrepare 1x PBSPBS is required to prepare reagents needed for this procedure. Dilute 10x PBS 1:10 in deionized water to make working 1x solution.", "[Reagent Preparation]\nPrepare a 4% paraformaldehyde solution.\nParaformaldehyde is toxic. Use gloves, wear a lab coat and work in a fume hood.", "...
null
null
null
dx.doi.org/10.17504/protocols.io.nradd2e
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
29,756
Enzymatic Assay of Trypsin Inhibition
null
dx.doi.org/10.17504/protocols.io.9a4h2gw
null
Neilier Junior
TITLE: Enzymatic Assay of Trypsin Inhibition AUTHORS: Neilier Junior [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol has been standardized for analysis of protease inhibitors in soybean leaf extract, but can be easily adjusted for other biological samples.</div></div> [STEPS] ?. [Prep...
["[Prepare the solutions and the workspace]\nPreparation of solutionsTrypsin solution:Dilute 1.25 mg of bovine trypsin in 1 mL of water pH 3.0 (adjust with HCl).L-BApNA stock solution:Dilute 130.47 mg L-BApNA in 5 mL DMSO (concentration: 6.0 10-2 M). Store at -20 °C and protected from light.L-BApNA usage solution (fres...
93,810
DNA Extraction from Inoculated Broth for WGS using the Maxwell RSC Cultured Cells DNA Kit
1
dx.doi.org/10.17504/protocols.io.6qpvr3z5zvmk/v1
https://www.protocols.io/view/dna-extraction-from-inoculated-broth-for-wgs-using-c7usznwe
Julie Haendiges, Ruth Timme
TITLE: DNA Extraction from Inoculated Broth for WGS using the Maxwell RSC Cultured Cells DNA Kit AUTHORS: Julie Haendiges, Ruth Timme [DESCRIPTION] The Maxwell® RSC Cultured cells DNA Kit is used with the Maxwell RSC or Maxwell RSC 48 Instrument from Promega. This extraction method purifies genomic DNA (gDNA) efficien...
["[Pre-processing of Gram Positive Cultures] Gram-Negative cultures do not require any steps prior to extraction on the Maxwell RSC instrument and can proceed to the Cultured Cells DNA Cartridge Preparation step. However, due to the thick peptidoglycan layer of Gram-Positive bacteria extra lysing is necessary. Follow t...
82,164
8. Taxon Group: Echinodermata
4
dx.doi.org/10.17504/protocols.io.4r3l27d63g1y/v2
https://www.protocols.io/view/8-taxon-group-echinodermata-cuguwtww
Kesella Scott-Somme, Chris Fletcher, Inez Januszczak
TITLE: 8. Taxon Group: Echinodermata AUTHORS: Kesella Scott-Somme, Chris Fletcher, Inez Januszczak [DESCRIPTION] This is part of the collection "DToL Taxon-specific Standard Operating Procedures (SOPs) for Marine Metazoa", lead by the Other Metazoa Working Group. The SOP collection contains guidance on how to process ...
[]
68,847
Genomic DNA isolation from fixed cells
1
dx.doi.org/10.17504/protocols.io.eq2lynm9qvx9/v1
https://www.protocols.io/view/genomic-dna-isolation-from-fixed-cells-cfgptjvn
Herschel Dhekne, Ebsy Jaimon, Suzanne R Pfeffer
TITLE: Genomic DNA isolation from fixed cells AUTHORS: Herschel Dhekne, Ebsy Jaimon, Suzanne R Pfeffer [DESCRIPTION] This protocol details the procedure of genomic DNA isolation from fixed cells. [STEPS] SECTION: Genomic DNA isolation from fixed cells 1. Resuspend 1-3 X106 cells in 250 µL of PBS and transfer to a 2 m...
["[Genomic DNA isolation from fixed cells] Resuspend 1-3 X106 cells in 250 µL of PBS and transfer to a 2 mL tube (this would represent one dish of 3T3 cells).", "[Genomic DNA isolation from fixed cells] Add 200 µg Proteinase K (from a 20 mg/mL stock) and 200 µg RNase A (from a 20 mg/mL stock).", "[Genomic DNA isolat...
null
null
null
dx.doi.org/10.17504/protocols.io.ci7uhm
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] How to make a 50 mg/ml Rifampicin Stock Solution [GUIDELINES] Make a 50 mg/ml Rifampicin Stock Solution [STEPS] ?. ?. ?.
[]
101,901
Sonication of α-synuclein Fibrils for injection into the mouse brain
1
dx.doi.org/10.17504/protocols.io.14egn337zl5d/v2
https://www.protocols.io/view/sonication-of-synuclein-fibrils-for-injection-int-dfrm3m46
Vijay Singh, Marta Castellana-Cruz, Nunilo Cremades, Laura Volpicelli-Daley
TITLE: Sonication of α-synuclein Fibrils for injection into the mouse brain AUTHORS: Vijay Singh, Marta Castellana-Cruz, Nunilo Cremades, Laura Volpicelli-Daley [DESCRIPTION] Animal models that accurately recapitulate the accumulation of alpha-synuclein (α-syn) inclusions, progressive neurodegeneration of the nigrost...
["[Sonicating Fibrils] Thaw and pipet 25-22 µL in 1.5 mL polystyrene sonication tube.", "[Sonicating Fibrils] WePFFs Fill Qsonica water reservoir with about 900 mL.", "[Sonicating Fibrils] Attach cooling system to Qsonica and set the temperature at 10 °C.", "[Sonicating Fibrils] Place 1.5 mL sonication tube with fibril...
82,896
CZ ID Workflow for Assembling Viral Consensus Genomes
1
dx.doi.org/10.17504/protocols.io.bp2l69ojklqe/v1
https://www.protocols.click/view/cz-id-workflow-for-assembling-viral-consensus-geno-cu7qwzmw
Karyna Rosario Cora, Elizabeth Fahsbender, CZ ID Team
TITLE: CZ ID Workflow for Assembling Viral Consensus Genomes AUTHORS: Karyna Rosario Cora, Elizabeth Fahsbender, CZ ID Team [DESCRIPTION] CZ ID's Viral Consensus Genome pipeline is designed to quickly assemble consensus genomes in bulk for any virus. Users can get started on their analysis by simply uploading Illumina...
["[Upload Data] Log in to your CZ ID account.", "[Upload Data] Navigate to the Upload page from the Discovery page by clicking the \"Upload\" link next to your username. Note that the upload process is divided into three general sections, including Samples, Metadata, and Review pages.", "[Upload Data] Select a project,...
60,311
Virtual Screening
6
dx.doi.org/10.17504/protocols.io.n92ldzr6ov5b/v1
https://www.protocols.io/view/virtual-screening-b65xrg7n
BOC Sciences
TITLE: Virtual Screening AUTHORS: BOC Sciences [DESCRIPTION] Virtual screening has been widely applied in early-stage drug discovery. As an alternative or complementary approach to high-throughput screening (HTS) assays with high cost and low hit rate, virtual screeningis an efficient computational method to identify...
[]
49,016
Grinding Tissue with the Qiagen Tissuelyzer (Soybean, Cowpea, Tobacco)
4
null
https://www.protocols.io/view/grinding-tissue-with-the-qiagen-tissuelyzer-soybea-bt4ynqxw
Steven Burgess, Lynn Doran
TITLE: Grinding Tissue with the Qiagen Tissuelyzer (Soybean, Cowpea, Tobacco) AUTHORS: Steven Burgess, Lynn Doran [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Grinding tissue using the Qiagen Tissuelyzer for DNA extraction, RNA extraction, or Immunoblotting.</div><div class = "text-block"><span s...
["Pre-cool components for tissue lysis.", "Pre-cool and sterilize 4mm SPEX stainless steel grinding beads in 100% ethanol in a beaker at for\n-80 °C", "Pre-cool the empty TissueLyser inserts for LN2 for by lowering the inserts into a full liquid nitrogen dewar with the extra-long tongs.\nWear lab coat, goggles, cryo...
11,410
Electrocompetent Agrobacterium preparation
null
dx.doi.org/10.17504/protocols.io.pdsdi6e
null
Magdalena Julkowska
TITLE: Electrocompetent Agrobacterium preparation AUTHORS: Magdalena Julkowska [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Preparing electrocompetent Agrobacterium - used for the GV301 strain that we currently have in stock in Salt Lab</div></div> [STEPS] ?. Grow seed culture to saturation (36-...
["Grow seed culture to saturation (36-48 hours), you will need ~5mL", "Inoculate 1 L of YEP media (20g peptone, 10 g yeast/ 1L) 1:200 with saturated agro. Alternatively inoculate 500ml YEP 1:100. Grow culture to an OD600 of 1.5 (24 hours or so). If you dont have YEP, just use normal LB media. Proceed on ice.", "Spin c...
103,202
Mouse brain slice electrophysiology
0
dx.doi.org/10.17504/protocols.io.rm7vzj7y8lx1/v1
https://www.protocols.io/view/mouse-brain-slice-electrophysiology-dg2a3yae
Shiyi Wang
TITLE: Mouse brain slice electrophysiology AUTHORS: Shiyi Wang [DESCRIPTION] Mouse brain slice electrophysiology [STEPS] 1. **Animal Handling** 1.1. - Anesthetize 3-4 mice of each genotype (WT and LRRK2 G2019Ski/ki) and condition using 200 mg/kg tribromoethanol (Avertin). 1.2. - Ensure proper anesthesia depth and con...
["**Animal Handling**", "- Anesthetize 3-4 mice of each genotype (WT and LRRK2 G2019Ski/ki) and condition using 200 mg/kg tribromoethanol (Avertin).", "- Ensure proper anesthesia depth and confirm absence of reflexes before proceeding.", "**Brain Extraction and Slicing**", "- Decapitate the mice and immediately immerse...
84,921
Feline Respiratory Pathogen Detection Assays
1
dx.doi.org/10.17504/protocols.io.ewov1q4z2gr2/v1
https://www.protocols.io/view/feline-respiratory-pathogen-detection-assays-cw6zxhf6
Come J Thieulent, Mariano Carossino, Laura Peak, Udeni B. R. Balasuriya
TITLE: Feline Respiratory Pathogen Detection Assays AUTHORS: Come J Thieulent, Mariano Carossino, Laura Peak, Udeni B. R. Balasuriya [DESCRIPTION] The Feline Respiratory Pathogen (FRP) Detection Assays is intended as an in vitro veterinary reagent set, based on quantitative PCR (qPCR) and Reverse Transcription qPCR (R...
["Thaw all reagents on ice.", "Centrifuge all reagents on a benchtop centrifuge to ensure no liquid is in cap and keep on ice", "Setup the Master Mix according to the following table 1:", "[PROGRAMMING THE THERMOCYCLER] Select the following fluorescence channels: ABYTM, Cy5, FAMTM, and VICTM.", "[PROGRAMMING THE THERMO...
93,454
Systematic Perturb-seq to discover endothelial cell programs related to CAD
4
dx.doi.org/10.17504/protocols.io.261ged73yv47/v1
https://www.protocols.io/view/systematic-perturb-seq-to-discover-endothelial-cel-c7hnzj5e
Gavin R. Schnitzler, Helen Kang, Glen Munson, Philine Guckelberger, Drew Bergman, Brian Cleary, Eric S Lander, Jesse Engreitz, Rajat M. Gupta
TITLE: Systematic Perturb-seq to discover endothelial cell programs related to CAD AUTHORS: Gavin R. Schnitzler, Helen Kang, Glen Munson, Philine Guckelberger, Drew Bergman, Brian Cleary, Eric S Lander, Jesse Engreitz, Rajat M. Gupta [DESCRIPTION] Linking variants from genome-wide association studies (GWAS) to underly...
["[General considerations for culturing of TeloHAEC] Telomerase-immortalized human aortic endothelial cells (TeloHAEC) were purchased from ATCC (CRL-4052), and grown in Lifeline VEGF endothelial cell media (LL-0005) with 1x Penn/Strep (ThermoFisher #15070063 or equivalent, used at 1:100 dilution) in place of the provid...
87,256
Lee et al. Digestion and Seeding (adapted from. Annika)
4
null
https://www.protocols.io/view/lee-et-al-digestion-and-seeding-adapted-from-annik-czfyx3pw
Annika Fendler
TITLE: Lee et al. Digestion and Seeding (adapted from. Annika) AUTHORS: Annika Fendler [DESCRIPTION] This Protocol is adapted from Lee et a. (2018) (https://www.sciencedirect.com/science/article/pii/S0092867418302976?via%3Dihub) The authors have established organoids from clusters of bladder cancer cells after TURB. ...
["[Prepare medium and enzyme mixes] Heat the shaking incubator to 37°C", "[Prepare medium and enzyme mixes] Prepare and adequate volume of Lee Organoid Medium and HBSS Washing Solution.", "[Thaw fragments]", "[Thaw fragments] Thaw 1 or more aliquots of fragments at 37 °C until only a small piece of ice remains", "[Prep...
36,003
Separating the Protein of Interest from MBP after TEV Protease Cleavage (NEB #E8201)
null
dx.doi.org/10.17504/protocols.io.bfebjjan
https://www.protocols.io/view/separating-the-protein-of-interest-from-mbp-after-bfebjjan
New England Biolabs
TITLE: Separating the Protein of Interest from MBP after TEV Protease Cleavage (NEB #E8201) AUTHORS: New England Biolabs [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><ul style = "list-style-type:disc;"><li style = "counter-reset:ol0;list-style-type:disc;"><span>Both the MBP-tag and TEV Protease ...
["[Resin Preparation]\nGently shake the Ni Resin bottle to completely resuspend the slurry and transfer desired amount to a closed column; allow resin to settle.", "[Resin Preparation]\nOpen column and allow storage buffer to flow through.", "[Resin Preparation]\nEquilibrate the resin by adding 5 column volumes (CV) of...
39,234
Performing CUT&RUN on adherent cells in a multi-well cell culture plate
4
dx.doi.org/10.17504/protocols.io.bijakcie
https://www.protocols.io/view/performing-cut-amp-run-on-adherent-cells-in-a-mult-bijakcie
Michi Miura
TITLE: Performing CUT&RUN on adherent cells in a multi-well cell culture plate AUTHORS: Michi Miura [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol describes a variant of the standard CUT&RUN procedure (Skene et al. </span><span style = "font-style:italic;">eLife</span><span> 201...
["[Cell seeding]\nSeed adherent cells in a standard 24-well cell culture plateSeed the cells at an appropriate density so that the cells are 70 to 90% confluent when performing CUT&RUN.", "[CUT&RUN]\nPerform CUT&RUNBefore cell permeabilisation (step 2.2),Remove the cell culture mediumWash the cells with PBS (Optional) ...
28,417
17 Inhibition Kinetics Measurement
null
dx.doi.org/10.17504/protocols.io.7y9hpz6
null
TJUSLS China
TITLE: 17 Inhibition Kinetics Measurement AUTHORS: TJUSLS China [STEPS] ?. [IC50 in vitro test] Soak the 96-well plates in 75% ethanol and put the container in ultrasonic cleaner for 30min to 1 hour, then use ddH2O to wash these plates several times. Put these clean plates in drying oven at 55°C. ?. [IC50 in vitro tes...
["[IC50 in vitro test]\nSoak the 96-well plates in 75% ethanol and put the container in ultrasonic cleaner for 30min to 1 hour, then use ddH2O to wash these plates several times. Put these clean plates in drying oven at 55°C.", "[IC50 in vitro test]\nDilute inhibitors using 100%DMSO to 8mM (a.k.a. 80μM in reaction syst...
21,152
UC Davis - RadioImmuno Assay (RIA) Protocol
null
dx.doi.org/10.17504/protocols.io.yv8fw9w
null
Peter Havel
TITLE: UC Davis - RadioImmuno Assay (RIA) Protocol AUTHORS: Peter Havel [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">The RIA (RadioImmuno Assay) is an assay method used for the quantification of various proteins. A ...
["Follow kit instructions for preparing standards and all reagents.", "Follow kit instructions for volumes and incubation times. Pipet standards and samples . Pipet antibody and tracer. Incubate.", "Follow kit instructions for volumes and incubation times. Add precipitating reagent. Incubate.", "Centrifuge.", "Aspirate...
30,700
Anti-DYKDDDDK Tag (L5) Affinity Gel Protocol
null
dx.doi.org/10.17504/protocols.io.98kh9uw
null
Sam Li
TITLE: Anti-DYKDDDDK Tag (L5) Affinity Gel Protocol AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Part I. Cell Lysate Preparation - A. Purifying DYKDDDDK-tag fusion proteins from crude E.coli extracts] Grow the bacteria cells under the condition that induce production of DYKDDDDK-tag fus...
["[Part I. Cell Lysate Preparation - A. Purifying DYKDDDDK-tag fusion proteins from crude E.coli extracts]\nGrow the bacteria cells under the condition that induce production of DYKDDDDK-tag fusion proteins.", "[Part I. Cell Lysate Preparation - A. Purifying DYKDDDDK-tag fusion proteins from crude E.coli extracts]\nHar...
35,930
Automated transfer of inactivated virus samples from individual 2ml tubes to 96-well plate
null
dx.doi.org/10.17504/protocols.io.bfb2jiqe
https://www.protocols.io/view/automated-transfer-of-inactivated-virus-samples-fr-bfb2jiqe
Efthymios Fidanis, Sophia Ward, Laura Cubitt, Amelia Edwards, Debbie Hughes, Michael Hubank, Jerome Nicod
TITLE: Automated transfer of inactivated virus samples from individual 2ml tubes to 96-well plate AUTHORS: Efthymios Fidanis, Sophia Ward, Laura Cubitt, Amelia Edwards, Debbie Hughes, Michael Hubank, Jerome Nicod [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Purpo...
["[Transfer of inactivated virus to 96 well plates]\nSample tubes will arrive from the CL2/3 facility after virus inactivation at . Working in the reception area (in a separate room), wipe outside of tube with alcohol wipe, invert tube twice, and place in centrifuge. Pulse spin tubes.\n0 Room temperature", "[Transfer o...
106,252
Immunofluorescence for pSer129 alpha-synuclein and total alpha-synuclein in mouse brain 
0
dx.doi.org/10.17504/protocols.io.5jyl82819l2w/v1
https://www.protocols.io/view/immunofluorescence-for-pser129-alpha-synuclein-and-djzk4p4w
Laura Volpicelli-Daley, mamenard
TITLE: Immunofluorescence for pSer129 alpha-synuclein and total alpha-synuclein in mouse brain  AUTHORS: Laura Volpicelli-Daley, mamenard [DESCRIPTION] This protocol details the immunofluorescence of brain section for phopho-serine129-alpha-synuclein and total alpha-synuclein [STEPS] SECTION: Day 1 1. Pre-warm sodium...
["[Day 1] Pre-warm sodium citrate buffer to 37 °C.", "[Day 1] Place sections in netwell inserts (Fisher Scientific 07-200-213) in 6-well plate.", "[Day 1] Wash 3X in cold 1X TBS for 5 min each with shaking (around 2 mL per well).", "[Day 1] Incubate at 37 °C for 60 min in pre-heated sodium citrate buffer (about 2 mL/we...
58,865
Plate Protein Expression on Autoinduction media
4
dx.doi.org/10.17504/protocols.io.b5qrq5v6
https://www.protocols.io/view/plate-protein-expression-on-autoinduction-media-b5qrq5v6
Stephane Fadanka, Chiara Gandini
TITLE: Plate Protein Expression on Autoinduction media AUTHORS: Stephane Fadanka, Chiara Gandini [DESCRIPTION] The current protocol describes the preparation and use of 2X YT autoinduction medium for recombinant protein expression on Petri dishes. This protocol allows for reproducible and time effective expression ex...
["[Preparation of the Overnight Pre-inoculum] Grow culture 10 mL of desired bacteria strain in LB broth supplemented with the appropriate antibiotic, grow overnight at 37 °C in an incubator.", "[Preparation of Auto-induction media] Prepare the needed amount of 4xYT autoinduction agar medium \n\n(It is recommended to p...
42,698
Thin Layer Chromatography (TLC) Bioautography
1
dx.doi.org/10.17504/protocols.io.bmxik7ke
https://www.protocols.io/view/thin-layer-chromatography-tlc-bioautography-bmxik7ke
Dane Lyddiard
TITLE: Thin Layer Chromatography (TLC) Bioautography AUTHORS: Dane Lyddiard [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A TLC bioautography protocol to test the constituents of plant extracts and essential oils for antibacterial activity was adapted and optimised from the methods described in Sm...
["Grow Bacteria (Overnight)Prepare streak plates of bacteria on appropriate solid agar media (e.g., nutrient or Mueller-Hinton agars). Incubate overnight (aerobically) at 35°C ±1°C", "Select TLC Solvent SystemCut TLC plates (handle with gloves) to 9 x 9 cm (or smaller for testing solvent systems). Using a TLC spotter o...
89,018
Preparing Cholesterol for use in NGM
1
dx.doi.org/10.17504/protocols.io.36wgq3oe5lk5/v1
https://www.protocols.io/view/preparing-cholesterol-for-use-in-ngm-c262yhge
Bonnie Evans
TITLE: Preparing Cholesterol for use in NGM AUTHORS: Bonnie Evans [DESCRIPTION] C. elegans requires cholesterol to progress through the four larval stages. [STEPS] 1. Prepare 5 mg/ml in Ethanol by resuspending the whole 250mg vial in 50 ml ~100% ethanol. 2. Filter sterilise using vacuum pump. 3. Store at 4 degree...
["Prepare 5 mg/ml in Ethanol by resuspending the whole 250mg vial in 50 ml ~100% ethanol.", "Filter sterilise using vacuum pump.", "Store at 4 degrees protected from the light"]
27,589
Protocol- Romantic relationships and family in Spaniards emerging adults
null
dx.doi.org/10.17504/protocols.io.67dhhi6
null
Marta Díez
TITLE: Protocol- Romantic relationships and family in Spaniards emerging adults AUTHORS: Marta Díez [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The exploration of and search for romantic relationships is one of the developmental tasks that characterize emerging adulthood, a new developmental ph...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.e46bgze
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>SWIFT™ Western Transfer Pads For Efficient High &amp; Low MW Protein Transfer</p> <p>(Cat. # <a href="http://www.gbiosciences.com/ResearchProducts/swift-desc.aspx" target="_blank">786‐370 to 786‐375, 786‐370S, 786‐373S</a>)</p> [BEFORE_START] <p>Combine 50ml of supplied SWIF...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ijpccmn
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>Adapted from Wetzel and Likens 2000. Limnological Analyses, Springer NY</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?.
[]
75,316
MDL-LDA Agile Method
1
dx.doi.org/10.17504/protocols.io.n2bvj82mxgk5/v2
https://www.protocols.io/view/mdl-lda-agile-method-cmsuu6ew
Freddie Prianes
TITLE: MDL-LDA Agile Method AUTHORS: Freddie Prianes [DESCRIPTION] As a guide in modeling the document labels using Latent Dirichlet allocation for archived documents in an Integrated Quality Assurance System (IQAS), the researchers used the agile method as it promotes flexibility, speed, and, most importantly, contin...
["[MDL-LDA Methodology] Plan: the researchers collected previous documents involved in the accreditation process, such as compliance reports under the areas of student, faculty, facility, library, and administration. Also, understanding of the existing problems in tracking, tagging, and duplication of these documents d...
79,602
Collecting needle and branch samples for terpenoid and expression analysis
1
null
https://www.protocols.io/view/collecting-needle-and-branch-samples-for-terpenoid-crysv7we
Vidya S Vuruputoor, Victoria Burton, Jill Wegrzyn
TITLE: Collecting needle and branch samples for terpenoid and expression analysis AUTHORS: Vidya S Vuruputoor, Victoria Burton, Jill Wegrzyn [DESCRIPTION] This protocol is used by the PCG group for sampling needle and branch tissue from hemlock conifers for terpenoid analysis, as well as RNA expression analysis. [STE...
["Open Treesnap (https://treesnap.org/), record the accession number on the tree and fill in notes for hemlock woolly adelgid infestation (HWA) levels (scale provided by the app).", "Use the comment field to enter: EHS present OR EHS absent to record the hemlock scale.", "Take at least 3 photos with TreeSnap. This incl...
94,452
Propionylation and tryptic digestion
1
dx.doi.org/10.17504/protocols.io.5jyl8pz36g2w/v1
https://www.protocols.io/view/propionylation-and-tryptic-digestion-c8guztww
Laura Corveleyn, Sigrid Verhelst
TITLE: Propionylation and tryptic digestion AUTHORS: Laura Corveleyn, Sigrid Verhelst [DESCRIPTION] Protocol to derivatize (propionylation) and digest histone proteins into peptides with the purpose of bottom-up label free LC-MS/MS analysis. In this method, aspecific overpropionylation at serine (S), threonine (T) and...
["[First propionylation] Start with vacuum dried sample (20 µg/sample)", "[First propionylation] Add 20 µL 1M triethylammonium bicarbonate (TEAB)", "[First propionylation] Add 20 µL prop-reagent (Isopropylalcohol:propionic anhydride (79:1))", "[First propionylation] Spin down & Incubate at room temperature for 30 minut...
38,945
Investigation of ancestral alleles in the Bovinae subfamily
5
dx.doi.org/10.17504/protocols.io.bh99j996
https://www.protocols.io/view/investigation-of-ancestral-alleles-in-the-bovinae-bh99j996
Maulana Naji, Yuri Utsunomiya, Johann Sölkner, Benjamin Rosen, Gábor Mészáros
TITLE: Investigation of ancestral alleles in the Bovinae subfamily AUTHORS: Maulana Naji, Yuri Utsunomiya, Johann Sölkner, Benjamin Rosen, Gábor Mészáros [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">In evolutionary theory, divergence and speciation can arise from long periods of reproductive isol...
["[Reads alignment and variant calling on individuals separately]\nDownload Read Sequences in format of SRA FASTQ", "[Reads alignment and variant calling on individuals separately]\nMapping with BWA-Mem", "[Reads alignment and variant calling on individuals separately]\nIndividual Haplotype Calling", "[Joint analysis o...
108,999
Effects of Intraoperative Hypothermia on Patients Undergoing Laparoscopic Surgery: A Retrospective Cohort Study
0
dx.doi.org/10.17504/protocols.io.14egn6d2yl5d/v1
https://www.protocols.io/view/effects-of-intraoperative-hypothermia-on-patients-dnpf5djn
maliwan oofuvong
TITLE: Effects of Intraoperative Hypothermia on Patients Undergoing Laparoscopic Surgery: A Retrospective Cohort Study AUTHORS: maliwan oofuvong [DESCRIPTION] This study aims to explore the association between intraoperative hypothermia and outcomes in adult patients undergoing laparoscopic surgery. A retrospective an...
["[Methodology] This study was approved by the Institutional Ethics Committee of the Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand in 11 July 2022 (REC 65-226-8-9). Since this is a retrospective study, informed consent was waived by the Institutional Ethics Committee of the Faculty of Medicine, ...
49,940
OMS Atlas FFPE Spatial Mapping
1
dx.doi.org/10.17504/protocols.io.buzunx6w
https://www.protocols.io/view/oms-atlas-ffpe-spatial-mapping-buzunx6w
Brett Johnson, Danielle Galipeau, Todd Camp, George Thomas
TITLE: OMS Atlas FFPE 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 a FFPE block, prepares the resulting slides, and then distributes t...
["[Preparation]\nVerify the identity of the FFPE block to be cut against written request for sectioning.", "[Preparation]\nLabel each Tanner slide with a unique BEMS ID and slide number, corresponding to the written request and FFPE spatial map (below).Note: Slides #1-4 compose the official OMS Atlas Dataset. ABC1Slid...
64,426
Fluxactive Complete Reviews – What are the Customers Saying?
3
dx.doi.org/10.17504/protocols.io.rm7vzyqp5lx1/v1
https://www.protocols.io/view/fluxactive-complete-reviews-what-are-the-customers-ca6ishce
healthy
TITLE: Fluxactive Complete Reviews – What are the Customers Saying? AUTHORS: healthy [DESCRIPTION] Fluxactive Complete has been released as a prostate wellness supplement that can support prostate health in men, which in turn helps keep their bladders and reproductive system healthy. [STEPS]
[]
92,381
Protocol for &#34;Development and characterization of a non-human primate model of diseeminated synucleinopathy&#34;
2
dx.doi.org/10.17504/protocols.io.81wgbxwnylpk/v1
https://www.protocols.io/view/protocol-for-34-development-and-characterization-o-c6f5zbq6
jlanciego, Alberto J. Rico, Julia Chocarro, Goiaz Ariznabarreta, Elvira Roda, Adriana Honrubia, Patricia Arnaiz, Almudena Corcho
TITLE: Protocol for &#34;Development and characterization of a non-human primate model of diseeminated synucleinopathy&#34; AUTHORS: jlanciego, Alberto J. Rico, Julia Chocarro, Goiaz Ariznabarreta, Elvira Roda, Adriana Honrubia, Patricia Arnaiz, Almudena Corcho [DESCRIPTION] Here we took advantage of a retrogradely-sp...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.hasb2ee
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol (like its companion protocol for liposome suspension and extrusion) was original created by <a href="&quot;http://www.whoi.edu/sbl/liteSite.do?litesite" target="_blank">Krista Longnecker</a> and <a href="http://jamesrco.github.io/" target="_blank">Jamie Collins<...
[]
95,838
Treatment and staining of iPSC-derived neurons for lysosomal phenotype analysis
0
dx.doi.org/10.17504/protocols.io.e6nvwdwedlmk/v1
https://www.protocols.io/view/treatment-and-staining-of-ipsc-derived-neurons-for-c9t6z6re
Jessica Chedid, Adahir Labrador-Garrido, Nicolas Dzamko
TITLE: Treatment and staining of iPSC-derived neurons for lysosomal phenotype analysis AUTHORS: Jessica Chedid, Adahir Labrador-Garrido, Nicolas Dzamko [DESCRIPTION] This protocol describes the preparation and treatment of neuronal cultures to be imaged for its analysis using the Opera Phenix high-content screening sy...
["[Live Cell experimental outline] 1. Prepare: DQ-red-BSA 1:100, Cytopainter green cell proliferation reagent 1:500 and Hoechst 1:100 in complete cell culture media. \nAlternatively prepare: Mitotracker 1:10.000, Lysosomal staining 1:500, Cytopainter 1:500 and Hoechst 1:100 in complete cell culture media. \n\n2. Gently...
49,208
Multiplexed, single-cell profiling of histone modifications with SCEPTRE
4
dx.doi.org/10.17504/protocols.io.buaynsfw
https://www.protocols.io/view/multiplexed-single-cell-profiling-of-histone-modif-buaynsfw
Marcus A Woodworth, Hao Yuan Kueh, Joshua C. Vaughan
TITLE: Multiplexed, single-cell profiling of histone modifications with SCEPTRE AUTHORS: Marcus A Woodworth, Hao Yuan Kueh, Joshua C. Vaughan [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Genome architecture and activity are tightly regulated during an organism’s development and function. Histone...
["[Cell Culture and Fixation]\nWash three times with of 1× PBS.\n250 µl", "[Immunofluorescence]\nWash cells three times with of 1× PBS.\n250 µl", "[Immunofluorescence]\nIncubate in of blocking solution for at .\n250 µl\n0 Room temperature", "[Immunofluorescence]\nIncubate in of primary solution at .\n250 µl...
26,103
Infant formula sample preparation for element analysis using PIXE/RBS
null
dx.doi.org/10.17504/protocols.io.5qxg5xn
null
Seth H. Frisbie, Erika J. Mitchell, Stéphane Roudeau, Florelle Domart, Asuncion Carmona, Richard Orterga
TITLE: Infant formula sample preparation for element analysis using PIXE/RBS AUTHORS: Seth H. Frisbie, Erika J. Mitchell, Stéphane Roudeau, Florelle Domart, Asuncion Carmona, Richard Orterga [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol was developed to quantify chemical elements fro...
["Work under a clean atmosphere to avoid dust contamination.", "Use a disposable plastic spatula to deliver between 1.0000 grams (g) and 1.0099 g of powdered product to a laboratory-grade plastic centrifuge tube.", "Add 10.00 milliliters (mL) of ultra-trace elemental analysis grade water (Water OptimaTM).", "Vortex unt...
73,112
A new metabarcoding approach to survey diversity at the species level of Arcellinida (Amoebozoa: Tubulinea)
1
dx.doi.org/10.17504/protocols.io.yxmvm2389g3p/v1
https://www.protocols.click/view/a-new-metabarcoding-approach-to-survey-diversity-a-cjmyuk7w
R. González-Miguéns
TITLE: A new metabarcoding approach to survey diversity at the species level of Arcellinida (Amoebozoa: Tubulinea) AUTHORS: R. González-Miguéns [DESCRIPTION] PCR protocol for the paper:" A needle in a haystack: a new metabarcoding approach to survey diversity at the species level of Arcellinida (Amoebozoa: Tubulinea)"...
["[Polymerase Chain Reaction protocol] First PCR\n\nPrimers:\nPrimer forward: LCO 1490 (5´ GGTCAACAAATCATAAAGATATTGG 3´)\nPrimer reverse: HCO 2198 (5´ TAAACTTCAGGGTGACCAAAAAATCA 3´)\n\nReagents for the PCR:\n \nTotal reaction mix per sample= 10 μL\n\n\nThermocycler program:\n \n\nDenaturation temperatures and times may...
79,877
Virus production
1
dx.doi.org/10.17504/protocols.io.8epv5jpkjl1b/v1
https://www.protocols.io/view/virus-production-cr9dv926
michela.deleidi
TITLE: Virus production AUTHORS: michela.deleidi [DESCRIPTION] Lentiviral production protocol [STEPS] SECTION: Virus production 2. HEK 293 T cells were seeded in a 10-cm dish to reach 80% confluency the day after transfection.The next day, the medium was changed. SECTION: Virus production 3. On days 4 and 5 after tra...
["[Virus production] HEK 293 T cells were seeded in a 10-cm dish to reach 80% confluency the day after\ntransfection.The next day, the medium was changed.", "[Virus production] On days 4 and 5 after transfection, the medium was collected and filtered through a0.45-\nμm PVDF membrane.", "[Virus production] For lentivira...
61,999
Annexin V/PI- Assay: Flow Based Medium Throughput Assessment of Apoptotic Response to Two and Three Drug Combinations
4
dx.doi.org/10.17504/protocols.io.14egn7my6v5d/v1
https://www.protocols.io/view/annexin-v-pi-assay-flow-based-medium-throughput-as-b8sprwdn
Dennis Juarez, dfruman
TITLE: Annexin V/PI- Assay: Flow Based Medium Throughput Assessment of Apoptotic Response to Two and Three Drug Combinations AUTHORS: Dennis Juarez, dfruman [DESCRIPTION] Basic principals and protocol for medium throughput cell viability screening in suspension cells using Annexin V/PI staining on the Attune NXT used...
["[Basic Principals and Planning Before Proceeding] Cells in culture can only tolerate small amounts of the vehicle in which a drug is dissolved in.\n\nAs such, you should minimize and standardize the amount of vehicle used in an experiment across all treatments.", "[Basic Principals and Planning Before Proceeding] I c...
null
null
null
dx.doi.org/10.17504/protocols.io.rv5d686
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
35,631
DNA and RNA stabilization
null
dx.doi.org/10.17504/protocols.io.be2pjgdn
https://www.protocols.io/view/dna-and-rna-stabilization-be2pjgdn
Neilier Junior
TITLE: DNA and RNA stabilization AUTHORS: Neilier Junior [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">DNA and RNA stabilization solution is an aqueous and non-toxic tissue storage reagent that rapidly permeates tissues to stabilize and protect nucleic acids in fresh specimens. Tissue pieces are h...
["[Stabilization solution preparation (1.5 L)]\n935 mL of autoclaved MilliQ water700 g ammonium sulfateStir until dissolved Add 25 mL of 1 M sodium citrateAdd 40 mL of 0.5 M EDTAAdjust to pH 5.2 using concentrated H2SO4Complete the volume to 1.5 L with autoclaved MilliQ water700 g ammonium sulfateStir until dissolved A...
80,199
&#34;Midnight&#34; SARS-CoV2 genome sequencing protocol using 1200bp amplicon primer set v2 and the Nanopore Rapid library kit
1
null
https://www.protocols.io/view/34-midnight-34-sars-cov2-genome-sequencing-protoc-csjfwcjn
Nikki Freed, Lydia Murphy, Benjamin Schwessinger, Olin Silander
TITLE: &#34;Midnight&#34; SARS-CoV2 genome sequencing protocol using 1200bp amplicon primer set v2 and the Nanopore Rapid library kit AUTHORS: Nikki Freed, Lydia Murphy, Benjamin Schwessinger, Olin Silander [DESCRIPTION] This protocols is part of the ANU Biosecurity mini-research project #2 "An SARS-COV2 incursion sc...
["[cDNA preparation] You will receive a PCR strip of eight with 6 tubes containing 2uL Lunascript RT supermix. Label the tubes with 1-6 and your RG name. Note down which sample corresponds to which number.\n\nMix the following components in an 0.2mL 8-strip tube on ice; \n\nComponent ...
49,790
Parzenie herbaty
1
null
https://www.protocols.io/view/parzenie-herbaty-buu6nwze
tomasz.zielinski
TITLE: Parzenie herbaty AUTHORS: tomasz.zielinski [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol is part of the lesson within the </span><a href="https://carpentries-incubator.github.io/fair-bio-practice/" style = "text-decoration:underline;color:blue;cursor:pointer;"><span sty...
["[Preparation of water and tea]\nNapełnij czajnik wodą, co najmniej 300ml wody.", "[Preparation of water and tea]\nW międzyczasie przygotuj herbatę, rozpakuj ją i umieść jedna torebkę w szklance. Pamiętaj wyłożyć etykietkę na zewnątrz szklanki.", "[Preparation of water and tea]\nJak tylko woda się zagotuje wlej szybko...
17,559
Comparison of two column agglutination tests for red blood cell antibody testing
null
dx.doi.org/10.17504/protocols.io.vdxe27n
null
Jonas Sawierucha, Marion Posset, Viola Hähnel, Christian L. Johnson, James A. Hutchinson, Norbert Ahrens
TITLE: Comparison of two column agglutination tests for red blood cell antibody testing AUTHORS: Jonas Sawierucha, Marion Posset, Viola Hähnel, Christian L. Johnson, James A. Hutchinson, Norbert Ahrens [STEPS] ?. [Dataset]
["[Dataset]"]
78,284
Synthetic Procedure of 3,3&#39;-dimethoxy-5,5&#39;-dipropyl-2,2&#39;-biphenyldiol
6
dx.doi.org/10.17504/protocols.io.yxmvm2mr6g3p/v1
https://www.protocols.io/view/synthetic-procedure-of-3-3-39-dimethoxy-5-5-39-dip-cqpkvvkw
Lisa.Stanley, rui.katahira, Gregg T. Beckham
TITLE: Synthetic Procedure of 3,3&#39;-dimethoxy-5,5&#39;-dipropyl-2,2&#39;-biphenyldiol AUTHORS: Lisa.Stanley, rui.katahira, Gregg T. Beckham [DESCRIPTION] A direct understanding of the degradation reaction pathways of lignin polymers in biomass is difficult due to the complexity of lignin’s structure. To overcome th...
["[Synthetic Procedure]", "[Synthetic Procedure] 4-propylguaiacol (5.00 g , 0.03 mol) was dissolved in acetone (75 mL ) then diluted with 500 mL of acetic acid buffer (pH 5) [see Note 1]. Oxygen gas (O2) was bubbled through the buffer solution for 30 minutes. 75 mg of laccase (>0.5 U/mg) was then added to the solution....
15,876
Legume-rhizobia interactions in a complex microbiome
null
dx.doi.org/10.17504/protocols.io.trcem2w
null
Ming-Dao Chia
TITLE: Legume-rhizobia interactions in a complex microbiome AUTHORS: Ming-Dao Chia [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Contains all documented protocols used for Ming-Dao Chia's Honours project, "Legume-rhizobia interactions in a complex microbiome"</div></div> [STEPS]
[]
62,100
Superior Keto: KETOGENIC DIET INGREDIENTS, BENEFITS AND POSSIBLE SIDE EFFECTS?
3
dx.doi.org/10.17504/protocols.io.bp2l6136rvqe/v1
https://www.protocols.io/view/superior-keto-ketogenic-diet-ingredients-benefits-b8vurw6w
Superior Keto
TITLE: Superior Keto: KETOGENIC DIET INGREDIENTS, BENEFITS AND POSSIBLE SIDE EFFECTS? AUTHORS: Superior Keto [DESCRIPTION] Superior Keto [STEPS]
[]