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59,242
Stranded Mapping from Oriented Long Reads
1
dx.doi.org/10.17504/protocols.io.b54iq8ue
https://www.protocols.io/view/stranded-mapping-from-oriented-long-reads-b54iq8ue
David A Eccles
TITLE: Stranded Mapping from Oriented Long Reads AUTHORS: David A Eccles [DESCRIPTION] This protocol demonstrates how to map strand-oriented long reads to a genome, and visualise them in a genome browser. The general idea is to use minimap2 to create stranded BAM files, which are split for forward/reverse orientatio...
["[Orient Reads] Orient reads as per protocol Preparing Reads for Stranded Mapping.\n\nIf this has been done, then the following command should produce output without errors:\n \nExample output:", "[Index Preparation] Prepare genome index for spliced alignment", "[Read Mapping] Map the long reads to the genome using mi...
null
null
null
dx.doi.org/10.17504/protocols.io.kcfcstn
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
55,776
MAVRICS: A Robust and Safe Magnetic Nanoparticle based RNA Extraction Method Compatible with Phenol-chloroform Inactivated Infectious Samples
1
dx.doi.org/10.17504/protocols.io.b2p8qdrw
https://www.protocols.io/view/mavrics-a-robust-and-safe-magnetic-nanoparticle-ba-b2p8qdrw
Mo Li, Gerardo Ramos-Mandujano
TITLE: MAVRICS: A Robust and Safe Magnetic Nanoparticle based RNA Extraction Method Compatible with Phenol-chloroform Inactivated Infectious Samples AUTHORS: Mo Li, Gerardo Ramos-Mandujano [DESCRIPTION] Diagnosis and surveillance of emerging pathogens such as SARS-CoV-2 depend on nucleic acid isolation from clinica...
["Silica magnetic nanoparticles (SiMNP) synthesis.\nSiMNP synthesis was done following the published protocols in BOMB.bio: BOMB magnetic core nanoparticles synthesis and BOMB coating ferrite MNPs with silica oxide.", "COVID-19 patient samples.\nOropharyngeal or nasopharyngeal swabs are steeped in 1 mL acid guanidinium...
24,090
Systemic analysis of injection site pain caused by subcutaneous administration of FKB327 and reference product via different delivery methods
null
dx.doi.org/10.17504/protocols.io.3r2gm8e
null
Takahiro Ito, Masayuki Takanuma, Yasumasa Arai
TITLE: Systemic analysis of injection site pain caused by subcutaneous administration of FKB327 and reference product via different delivery methods AUTHORS: Takahiro Ito, Masayuki Takanuma, Yasumasa Arai [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">FKB327 is a biosimilar of the adalimumab refere...
["Data to be analyzed in systemic analysisThe VAS data of injection site pain from FKB327-001, -002, -003, and -004 are integrated for the systemic analysis. All cumulative data will be analyzed for the randomized subjects with injection site pain data collected immediately after injection at Day 1. Some subjects had m...
null
null
null
dx.doi.org/10.17504/protocols.io.j3xcqpn
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
61,966
Chemically Labile Linkers
6
dx.doi.org/10.17504/protocols.io.36wgq7mexvk5/v1
https://www.protocols.io/view/chemically-labile-linkers-b8rnrv5e
BOC Sciences
TITLE: Chemically Labile Linkers AUTHORS: BOC Sciences [DESCRIPTION] The chemically labile linkers, which include acid-cleavable linkers and disulfide linkers, are extensively applied to ADCs due to their ability to undergo fracture, increasing the acidity of the endosomal-lysosomal pathway, as well as the concen...
[]
97,917
Filaggrin genotyping using Taqman SNP genotyping assays
0
dx.doi.org/10.17504/protocols.io.3byl492mogo5/v1
https://www.protocols.io/view/filaggrin-genotyping-using-taqman-snp-genotyping-a-dbu52ny6
Angie Fawkes, Katarzyna Hafezi, Lee Murphy
TITLE: Filaggrin genotyping using Taqman SNP genotyping assays AUTHORS: Angie Fawkes, Katarzyna Hafezi, Lee Murphy [DESCRIPTION] The FLG gene encodes the filaggrin protein which is essential for epidermal barrier formation and hydration. Mutations in the filaggrin gene are associated with a broad range of skin and all...
["[Prepare reaction mix] The tables below are for one sample based on 10μL reaction volume (2μL of DNA and 8μL of reaction mix). Multiply up reaction mix by number of samples plus extra for dead-volume. Ensure a sensible amount for accurate pipetting. \n(This means that for R501X and S3247X primers will be at 300nM & p...
33,927
Q5® Site-Directed Mutagenesis (E0554)
1
dx.doi.org/10.17504/protocols.io.bddfi23n
https://www.protocols.io/view/q5-site-directed-mutagenesis-e0554-bddfi23n
New England Biolabs
TITLE: Q5® Site-Directed Mutagenesis (E0554) AUTHORS: New England Biolabs [DESCRIPTION] This is the protocol for the Q5® Site-Directed Mutagenesis Kit (E0554). [GUIDELINES] DESCRIPTION The Q5® Site-Directed Mutagenesis Kit enables rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hour...
["[Exponential Amplification (PCR)] Assemble the following reagents in a thin-walled PCR tube.\n 25 μl RXN FINAL CONC. Q5 Hot Start High-Fidelity 2X Master Mix 12.5 μl 1X 10 μM Forward Primer 1.25 μl 0.5 μM 10 μM Reverse Primer 1.25 μl 0.5 μM Template DNA (1–25 ng/μl) 1 μl 1-25 ng Nuclease-free water 9.0 ...
53,375
DNA Ethanol Precipitation (SOP009.v1.1)
4
null
https://www.protocols.io/view/dna-ethanol-precipitation-sop009-v1-1-byc7pszn
Rory Kruithoff, Douglas Shepherd
TITLE: DNA Ethanol Precipitation (SOP009.v1.1) AUTHORS: Rory Kruithoff, Douglas Shepherd [DESCRIPTION] Document Summary: This document, SOP002 - DNA Ethanol Precipitation, describes a method to concentrate or dry DNA using ethanol to reduce the solubility of dissolved DNA causing it to precipitate out of solution. ...
["[Precipitate DNA] Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample.", "[Precipitate DNA] Mix, and store at -20 °C for at least 60 min to precipitate the DNA.  Typically, DNA will be left at -20 °C 60 minbefore proceeding to part 2.", "[Pellet DNA and dry] Reco...
31,453
Association between circulating microRNA-122, microRNA-126-3p and microRNA-146a and inflammatory markers in patients with pre-diabetes and type 2 diabetes mellitus
1
dx.doi.org/10.17504/protocols.io.bax5ifq6
https://www.protocols.io/view/association-between-circulating-microrna-122-micro-bax5ifq6
Hassan Mozaffari-Khosravi, Fahime Zeinali, Seyed Mohsen Aghaei Zarch, Alireza Jahan-mihan, Shima Abdollahi, Seyed Mehdi Kalantar, Mohammad Yahya Vahidi Mehrjardi, Hossein Fallahzadeh, Mahdieh Hosseinzadeh-Shamsi-Anar, Masoud Rahmanian
TITLE: Association between circulating microRNA-122, microRNA-126-3p and microRNA-146a and inflammatory markers in patients with pre-diabetes and type 2 diabetes mellitus AUTHORS: Hassan Mozaffari-Khosravi, Fahime Zeinali, Seyed Mohsen Aghaei Zarch, Alireza Jahan-mihan, Shima Abdollahi, Seyed Mehdi Kalantar, Mohammad Y...
["[Real time PCR]\nReal time PCR", "[IL6 measurement]\nIL6 measurement by ELISA kit https://www.diaclone.com/documents/protocole/950.030_Human_IL-6_ELISA_kit_insert_v11.pdf", "[RNA extraction]\nRNA extraction http://tribioscience.com/files/315-150.pdf", "[RNA extraction]\nPrepare 750 ul RiboExTM LS in a 1.5 ml microcen...
97,862
Nuclei Isolation for Human Ovary Explants
0
dx.doi.org/10.17504/protocols.io.e6nvw1632lmk/v1
https://www.protocols.io/view/nuclei-isolation-for-human-ovary-explants-dbte2nje
Nicolas Martin
TITLE: Nuclei Isolation for Human Ovary Explants AUTHORS: Nicolas Martin [DESCRIPTION] This protocol uses fresh frozen human ovary explants to isolate nuclei suspension. [GUIDELINES] This protocol needs prior approval by the users' institutional review board (IRB) or equivalent ethics committee(s). [STEPS] SECTION: ...
["[Nuclei Isolation Protocol for Human Ovary Explants] Chapter 1—Single Cell Gene Expression & Chromium Fixed RNA Profiling of the protocol CG000505 REV A was used to isolate nuclei from frozen human ovary explants with the following modifications: 1) a cordless motor pestle (VWR, Catalog number 47747-370) was used for...
20,451
iPSC PCR: For Screening Edited Clones
null
dx.doi.org/10.17504/protocols.io.x8bfrsn
null
Celeste Karch, Rita Martinez, Jacob Marsh
TITLE: iPSC PCR: For Screening Edited Clones AUTHORS: Celeste Karch, Rita Martinez, Jacob Marsh [STEPS] ?. [PCR] This is the set up for a 50 µL reaction. ABC1Volumex# rxns25x Green GoTaq Flexi Buffer10 µl325mM MgCl26 µl425mM dNTPs0.8 µl5Forward Primer (10uM)2 µl6Reverse Primer (10uM)2 µl7GoTaq DNA Polymerase (5U/uL)0...
["[PCR]\nThis is the set up for a 50 µL reaction. ABC1Volumex# rxns25x Green GoTaq Flexi Buffer10 µl325mM MgCl26 µl425mM dNTPs0.8 µl5Forward Primer (10uM)2 µl6Reverse Primer (10uM)2 µl7GoTaq DNA Polymerase (5U/uL)0.25 µl8Milli-Q H2026.95 µl9Quick Extract gDNA2 µl10Total50 µl\nABC1Volumex# rxns25x Green GoTaq Flexi Buf...
92,384
SOP for genome size estimations from collection specimens
3
dx.doi.org/10.17504/protocols.io.yxmvm3by9l3p/v1
https://www.protocols.io/view/sop-for-genome-size-estimations-from-collection-sp-c6f8zbrw
Jean François Flot, Mohammed Tawfeeq, Ana Riesgo
TITLE: SOP for genome size estimations from collection specimens AUTHORS: Jean François Flot, Mohammed Tawfeeq, Ana Riesgo [DESCRIPTION] Genome size data is crucial now for comparative studies with the goal of understanding the mechanisms of genomic change and their biological implications related to genome size diver...
[]
52,282
Growth rate determination of E. coli
4
dx.doi.org/10.17504/protocols.io.bxa2pige
https://www.protocols.io/view/growth-rate-determination-of-e-coli-bxa2pige
Ashwinuday
TITLE: Growth rate determination of E. coli AUTHORS: Ashwinuday [DESCRIPTION] <div class = "text-blocks"><br/><div class = "text-block"><span>This is to measure the growth rate of </span><span style = "font-style:italic;">E. coli.</span></div><br/><div class = "text-block">The initially used culture is a primary cult...
["Grow the bacteria in an overnight (14-18 hours) culture.", "Inoculate 50 ml autoclaved culture media in a conical flask with a part of the overnight grown culture equal to 2% volume of fresh media (i.e. 1ml) inside a laminar hood.", "Put the rest of the freshly inoculated flask into a shaking incubator at at .\n33\...
null
null
null
dx.doi.org/10.17504/protocols.io.f7cbriw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>prepare 100 mL per 250 mL flask for each overnight culture</p> <p>for     </p> <h1>CZV8CS - liquid media for flask culture</h1> <p>prepare 100 mL per 250 mL flask for each overnight culture for</p> <p>                                                   <strong><u>100 mL</u></s...
[]
94,242
Protocol3_HEK293T cells and mRNA transfections.pdf
1
dx.doi.org/10.17504/protocols.io.36wgq3zyklk5/v1
https://www.protocols.io/view/protocol3-hek293t-cells-and-mrna-transfections-pdf-c8aazsae
angelica.gonzalez
TITLE: Protocol3_HEK293T cells and mRNA transfections.pdf AUTHORS: angelica.gonzalez [DESCRIPTION] HEK293T cells and mRNA transfections [STEPS]
[]
90,197
Immunohistochemistry protocol optimized at iMM-JLA
1
dx.doi.org/10.17504/protocols.io.j8nlkon2wv5r/v1
https://www.protocols.io/view/immunohistochemistry-protocol-optimized-at-imm-jla-c4bvysn6
Ana M Biscaia Santos, Ana M Cristóvão Pinto, Ana R Pires, Joana G Antunes
TITLE: Immunohistochemistry protocol optimized at iMM-JLA AUTHORS: Ana M Biscaia Santos, Ana M Cristóvão Pinto, Ana R Pires, Joana G Antunes [DESCRIPTION] The immunohistochemistry (IHC) protocol is used to showcase the location of specific proteins in relation to the arquitecture of the tissue. In this method, we use...
["[Preparation of Wash Buffer] Measure 20 mL of 20X", "[Preparation of Wash Buffer] Make up with distilled water to 1 L of total volume", "[Antigen Retrieval] Perform antigen retrieval using PT Module by Thermo Scientific with a cycle of pre-heat to 65ºC and heat to 95ºC for 20 minutes.", "[Counterstain and mount] Clea...
67,557
Monkeypox virus multiplexed PCR amplicon sequencing (PrimalSeq) V.2. 
1
dx.doi.org/10.17504/protocols.io.5qpvob1nbl4o/v2
https://www.protocols.io/view/monkeypox-virus-multiplexed-pcr-amplicon-sequencin-cd8ds9s6
Nicholas F.G. Chen*, Luc Gagne*, Matthew Doucette, Sandra Smole, Erika Buzby, Joshua Hall, Stephanie Ash, Rachel Harrington, Seana Cofsky, Selina Clancy, Curtis J Kapsak, Joel Sevinsky, Kevin Libuit, mallery.breban , Chrispin Chaguza, Nathan D. Grubaugh, Daniel J. Park, Glen R. Gallagher#, Chantal B.F. Vogels#
TITLE: Monkeypox virus multiplexed PCR amplicon sequencing (PrimalSeq) V.2.  AUTHORS: Nicholas F.G. Chen*, Luc Gagne*, Matthew Doucette, Sandra Smole, Erika Buzby, Joshua Hall, Stephanie Ash, Rachel Harrington, Seana Cofsky, Selina Clancy, Curtis J Kapsak, Joel Sevinsky, Kevin Libuit, mallery.breban , Chrispin ...
["[Dilute and Pool Primers] Reagents:", "[Dilute and Pool Primers] If not already done, separate odd and even numbered primer pairs into two separate boxes. These will constitute the two pools", "[Dilute and Pool Primers] Label 164, 8-strip tubes with the corresponding odd-numbered primer name (e.g. 3 left)", "[Dilute ...
null
null
null
dx.doi.org/10.17504/protocols.io.ci4ugv
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] How to make a 1M DTT Stock Solution [STEPS] ?. ?. ?. ?.
[]
25,877
Permanent multiprofessional auditory assessment protocol for children with congenital zika virus syndrome
null
dx.doi.org/10.17504/protocols.io.5hvg366
null
Klinger Vagner Teixeira da Costa, Pedro Menezes, Ana Frizzo
TITLE: Permanent multiprofessional auditory assessment protocol for children with congenital zika virus syndrome AUTHORS: Klinger Vagner Teixeira da Costa, Pedro Menezes, Ana Frizzo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Trata-se de 0um protocolo de avaliação auditiva multiprofissional para...
[]
36,353
PCOS lifestyle program
null
dx.doi.org/10.17504/protocols.io.bfq9jmz6
https://www.protocols.io/view/pcos-lifestyle-program-bfq9jmz6
Geranne Jiskoot, J.S.E Laven, J.J. van Busschbach, C. de Klerk, J. de Niet
TITLE: PCOS lifestyle program AUTHORS: Geranne Jiskoot, J.S.E Laven, J.J. van Busschbach, C. de Klerk, J. de Niet [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Obesity in women with polycystic ovary syndrome (PCOS)</div><div class = "text-block">negatively affects all clinical features, and a 5 to...
["[Background]\nPolycystic ovary syndrome (PCOS) is a common endocrine disorder that affects 5–10% of women in their reproductive years [1]. According to the ESHRE/ASRM Rotterdam consensus [2], the diagnosis of PCOS requires at least 2 of the following 3 criteria: oligoovulation or anovulation (irregular or no menstrua...
63,000
Weight Crasher Keto Gummies
3
dx.doi.org/10.17504/protocols.io.bp2l61m8kvqe/v1
https://www.protocols.io/view/weight-crasher-keto-gummies-b9ryr57w
H Douglas Morris
TITLE: Weight Crasher Keto Gummies AUTHORS: H Douglas Morris [DESCRIPTION] Weight Crasher Keto Gummies is a natural enhancement proposed for weight reduction. These diet pills are intended for people who are now on the ketogenic diet and aim to improve their belongings. [STEPS]
[]
77,741
Calcium fluorimetry with the FLIPR Calcium 6 kit on FlexStation 3
1
dx.doi.org/10.17504/protocols.io.3byl4jdmjlo5/v1
https://www.protocols.io/view/calcium-fluorimetry-with-the-flipr-calcium-6-kit-o-cp6mvrc6
Angus Li
TITLE: Calcium fluorimetry with the FLIPR Calcium 6 kit on FlexStation 3 AUTHORS: Angus Li [DESCRIPTION] This protocol details an experimental procedure used to generate results described in the manuscript Li, A., Liu, S., Huang, R., Ahn, S., & Lefkowitz, R. J. (2023). Loss of biased signaling at a G protein-coupled r...
["Plate U2OS-TetOn-AT1R in microplates (Black with clear bottom, lysine- coated Corning 3842) at 15000 cells/well 2 days prior to assay", "Add doxycycline and optionally PTX to wells 14 hours before replacement with loading buffer", "Remove one vial of Calcium 6 Assay Reagent (Component A, good for 2 plates) from the f...
39,140
S30-S30A-S30-Buffers- Haseloff Lab
4
dx.doi.org/10.17504/protocols.io.bigckbsw
https://www.protocols.io/view/s30-s30a-s30-buffers-haseloff-lab-bigckbsw
Fernando Guzman Chavez, Jim Haseloff
TITLE: S30-S30A-S30-Buffers- Haseloff Lab AUTHORS: Fernando Guzman Chavez, Jim Haseloff [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:center"><span>Following this recipe, you will obtain 1L of</span><span style = "font-weight:bold;"> S30</span><span style...
["[1. S30 Buffer]\nTo prepare S30 buffer, following compounds have to be ready-to-use:Tris-AcetateMagnesium acetate tetrahydratePotassium acetate1M DTT5M KOH", "[1. S30 Buffer]\n1. 1 Prepare for autoclavingVolumes indicated are sufficient for 1 L of S30 buffer Before useadd 2 mL of 1 M DTT(2mM, final concentration)For ...
null
null
null
dx.doi.org/10.17504/protocols.io.cjjukm
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
85,936
Whole-cell proteomics and Analysis with or without nutrient stress by Tandem Mass Tagging-based proteomics V2
4
dx.doi.org/10.17504/protocols.io.yxmvm32nbl3p/v1
https://www.protocols.io/view/whole-cell-proteomics-and-analysis-with-or-without-cx6qxrdw
Sharan Swarup, J. Wade Harper wade_harper@hms.harvard.edu, Kelsey Hickey
TITLE: Whole-cell proteomics and Analysis with or without nutrient stress by Tandem Mass Tagging-based proteomics V2 AUTHORS: Sharan Swarup, J. Wade Harper wade_harper@hms.harvard.edu, Kelsey Hickey [DESCRIPTION] The analysis of relative protein abundance has emerged as an important tool in cell biology. Typically, it...
["[Harvest, precipitation and digestion] For whole proteome analysis, 50 µg is required for each replicate. Cells from step 2 are washed with PBS three times. Cells were lysed by in UREA denaturing buffer (8M Urea, 150mM NaCl, 50mM EPPS pH8.0, containing mammalian protease inhibitor cocktail (Sigma), and Phos-STOP)...
49,235
space_publications_versions protocol 1 v2
1
null
https://www.protocols.io/view/space-publications-versions-protocol-1-v2-bubtnsnn
Maria Guliakina
TITLE: space_publications_versions protocol 1 v2 AUTHORS: Maria Guliakina [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">test</div></div> [STEPS] ?. test ?. est 2 ?. ?. test 4
["test", "est 2", "test 4"]
63,866
Chemotaxonomy-based Drug Discovery
4
dx.doi.org/10.17504/protocols.io.36wgq7yw5vk5/v1
https://www.protocols.io/view/chemotaxonomy-based-drug-discovery-cak2scye
contact.microbialtec
TITLE: Chemotaxonomy-based Drug Discovery AUTHORS: contact.microbialtec [DESCRIPTION] With the large numbers of already known microbial and plant metabolites, one of the major challenges in modern natural product discovery is to detect already known and trivial compounds rapidly. This relies on information managemen...
[]
81,575
Protocol for t-DNA insertional mutagenesis in the chickpea Fusarium wilt pathogen, Fusarium oxysporum f. sp. ciceris for identification of virulence genes
4
null
https://www.protocols.io/view/protocol-for-t-dna-insertional-mutagenesis-in-the-ctwfwpbn
Ramawatar Nagar
TITLE: Protocol for t-DNA insertional mutagenesis in the chickpea Fusarium wilt pathogen, Fusarium oxysporum f. sp. ciceris for identification of virulence genes AUTHORS: Ramawatar Nagar [DESCRIPTION] Chickpea is the third most consumed grain legume in the world. It serves as a valuable source of protein and micronutr...
["Preparing Fusarium spore suspension", "A wilting isolate (on the genotype JG62) of the Foc was cultured on PDA plates containing cefotaxime (50mg/L) at 25 °C for 7-10 days", "Spores were collected from the plant by adding 4 mL of double-distilled autoclave water to the plate and gently brushing the mycelium with a cl...
null
null
null
dx.doi.org/10.17504/protocols.io.jgacjse
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <ol> <li><strong> DNA Extraction</strong></li> </ol> <ul> <li>Extract DNA from sample using the phenol/chloroform procedure or your kit of choice. We typically use the Mo Bio Power Soil DNA extraction kit for extracting DNA from soil and plant litter samples.</li> <li>Use the Na...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.qv8dw9w
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Adapted from Tara Oceans extraction protocol (see citation).</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
26,168
Make NGM medium
null
dx.doi.org/10.17504/protocols.io.5syg6fw
null
Cancer Research UK / Wellcome Trust Gurdon Institute media kitchen
TITLE: Make NGM medium AUTHORS: Cancer Research UK / Wellcome Trust Gurdon Institute media kitchen [STEPS] ?. ?. Add bacto peptone and NaCl ?. Measure approx 4.8L double distilled H2O in 5L bell jar with a magnetic flea. ?. Stir until all solutes are dissolved. ?. Dispense approx 972ml NGM media per 1L bottle OR 48...
["Add bacto peptone and NaCl", "Measure approx 4.8L double distilled H2O in 5L bell jar with a magnetic flea.", "Stir until all solutes are dissolved.", "Dispense approx 972ml NGM media per 1L bottle OR 486ml NGM into a 500ml bottle.", "Dispense 17g of agar and a magnetic flea to each 1L bottle OR 8.5g of agar and a ...
61,925
PNA/DNA Chimera Synthesis
6
dx.doi.org/10.17504/protocols.io.j8nlkkmnwl5r/v1
https://www.protocols.io/view/pna-dna-chimera-synthesis-b8qdrvs6
Cathy Miller
TITLE: PNA/DNA Chimera Synthesis AUTHORS: Cathy Miller [DESCRIPTION] Pure PNA has significant binding properties, but it has completely different properties from DNA and RNA. For example, it cannot be recognized as a substrate when it interacts with nucleic acid modifying enzymes. On the one hand, this is an advantag...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ibccaiw
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
102,752
HEP-TILE: HBV whole genome sequencing (nanopore protocol)
1
dx.doi.org/10.17504/protocols.io.5jyl82bedl2w/v1
https://www.protocols.io/view/hep-tile-hbv-whole-genome-sequencing-nanopore-prot-dgj83urw
Sheila Lumley, Chris Kent, Josh Quick, Philippa Matthews
TITLE: HEP-TILE: HBV whole genome sequencing (nanopore protocol) AUTHORS: Sheila Lumley, Chris Kent, Josh Quick, Philippa Matthews [DESCRIPTION] This protocol describes the HEP-TILE tiled amplicon protocol for whole genome sequencing of Hepatitis B virus (HBV) on the nanopore MinION. We developed a pan-genotypic (gen...
["[Multiplex PCR] Set up the two PCR reactions per sample as follows in strip-tubes or plates. Gently mix by pipetting and pulse spin the tube to collect liquid at the bottom of the tube.", "[Multiplex PCR] Add 2.5 µL cDNA to each of the PCR reactions, gently mix by pipetting and pulse spin the tube to collect liquid a...
58,678
Minimum Inhibitory Concentration
4
null
https://www.protocols.io/view/minimum-inhibitory-concentration-b5iwq4fe
Amanda T Alker, Nicholas J Shikuma
TITLE: Minimum Inhibitory Concentration AUTHORS: Amanda T Alker, Nicholas J Shikuma [DESCRIPTION] Protocol for determining the minimum inhibitory concentration of antibiotics for marine bacteria. [STEPS] SECTION: Day 1 1. Streak out marine microbes onto Marine Agar plates and incubate overnight at 25-28ºC. SECTI...
["[Day 1] Streak out marine microbes onto Marine Agar plates and incubate overnight at 25-28ºC.", "[Day 2] Inoculate a single colony into 5mL Marine Broth (2216) media in the late afternoon/ early evening. Incubate overnight at 25ºC, shaking at 200 rpm.", "[Day 3] Measure optical density (OD600) of overnight culture gr...
63,431
20 Reasons You Need to Stop Stressing About Prima Slimming Pills UK
1
dx.doi.org/10.17504/protocols.io.q26g746bqgwz/v1
https://www.protocols.io/view/20-reasons-you-need-to-stop-stressing-about-prima-b97fr9jn
buddymurphyz
TITLE: 20 Reasons You Need to Stop Stressing About Prima Slimming Pills UK AUTHORS: buddymurphyz [DESCRIPTION] Prima Slimming Pills UK : Offers! Prima Slimming Pills UK Obesity issues are growing rapidly in the world. If you will not take care of your health and will keep on eating unhealthy food regularly, then it ...
["[20 Reasons You Need to Stop Stressing About Prima Slimming Pills UK]"]
65,880
Golden Revive Plus Reviews: [Scam Exposed 2022] Read Ingredients, Side Effects Price &amp; Where To Buy?
3
dx.doi.org/10.17504/protocols.io.kxygxzx7kv8j/v1
https://www.protocols.io/view/golden-revive-plus-reviews-scam-exposed-2022-read-ccjysupw
G
TITLE: Golden Revive Plus Reviews: [Scam Exposed 2022] Read Ingredients, Side Effects Price &amp; Where To Buy? AUTHORS: G [DESCRIPTION] Golden Reviews Plus Reviews [STEPS]
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86,815
Deparaffinization for tissue and organoids
4
null
https://www.protocols.io/view/deparaffinization-for-tissue-and-organoids-cyz7xx9n
Gabriela Vallejo Flores, Annika Fendler
TITLE: Deparaffinization for tissue and organoids AUTHORS: Gabriela Vallejo Flores, Annika Fendler [DESCRIPTION] This protocol is to withdraw the paraffin from formalin-fixed paraffin-embedded tissue and organoids and to recover the protein structure of your sample. [BEFORE_START] Prepare the steamer and the cook ...
["[Antigen retrieval solution] Transfer the slide into the antigen retrieval solution in the steamer, turn on the cook heater and incubate 5 min after the valve is up, turn off the cook heater and wait till the valve get down again.", "[Antigen retrieval solution] Transfer the slides into destillated water, now the sli...
78,074
Preparation of Encoding Probes SOP005.v1.5 (PCR, In-vitro Transcription, Reverse Transcription and USER ENZYME Digest)
4
dx.doi.org/10.17504/protocols.io.kqdg36zxqg25/v3
https://www.protocols.io/view/preparation-of-encoding-probes-sop005-v1-5-pcr-in-cqg2vtye
Rory Kruithoff, Douglas Shepherd
TITLE: Preparation of Encoding Probes SOP005.v1.5 (PCR, In-vitro Transcription, Reverse Transcription and USER ENZYME Digest) AUTHORS: Rory Kruithoff, Douglas Shepherd [DESCRIPTION] Document Summary:This document, Preparation of Encoding Probes (SOP005), describes the proc...
["[Part 1 - PCR Amplification - Step 1: Prepare the PCR reaction] In a 1.7 mL Eppendorf tube, mix the following:\n- 40 µL ;\n-  2 µL ;\n-  2 µL ; \n- 1 µL ; \n-  355 µL ; \n-  400 µL .", "[Part 1 - PCR Amplification - Step 1: Prepare the PCR reaction] Aliquot 25 µL into 32 PCR tubes.", "[Part 1 - PCR Amplification - ...
null
null
null
dx.doi.org/10.17504/protocols.io.hjpb4mn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This is a variation of the CagA translocation assay performed in our lab.</p> <p>This protocol has been used in the publication DOI: <a href="https://doi.org/10.1111/cmi.12166" target="_blank">10.1111/cmi.12166</a> </p> <p> </p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
97,156
10x Protocols: Chromium Single Cell/Nuclei Gene Expression Flex Multiplex -- University of Minnesota TMCs (CG000527 Rev E)
1
dx.doi.org/10.17504/protocols.io.bp2l6x81klqe/v2
https://www.protocols.io/view/10x-protocols-chromium-single-cell-nuclei-gene-exp-da5c2g2w
IOx Genomics, Laura Niedernhofer, David A Bernlohr
TITLE: 10x Protocols: Chromium Single Cell/Nuclei Gene Expression Flex Multiplex -- University of Minnesota TMCs (CG000527 Rev E) AUTHORS: IOx Genomics, Laura Niedernhofer, David A Bernlohr [DESCRIPTION] 10x Genomics Chromium Single Cell Expression flex protocol for library construction. Note: These protocols may no...
["[Preparation] Complete single cell or nuclei isolation and 10x fixation prior to starting this protocol", "[Library Preparation]", "[FASTQ Generation] BCL data from Illumina sequencer is demultiplexed and converted into FASTQ format using bcl2fastq version 2.20.0"]
53,852
Batch extraction of morphological and color metrics from invertebrate samples.
1
dx.doi.org/10.17504/protocols.io.byt4pwqw
https://www.protocols.io/view/batch-extraction-of-morphological-and-color-metric-byt4pwqw
Michael D Weiser, Katie E. Marshall, Cameron D. Siler, Michael Kaspari
TITLE: Batch extraction of morphological and color metrics from invertebrate samples. AUTHORS: Michael D Weiser, Katie E. Marshall, Cameron D. Siler, Michael Kaspari [DESCRIPTION] This protocol is the complete methods used to extract abundance, morphology and color data from samples of invertebrates. We developed th...
["[Camera Setup] We use a Canon EOS 5Ds (Model DS526521) camera fitted with a Canon EF 35mm f/2 IS USM Lens (Model 9523B002). To avoid having to recharge the battery we use a fixed power supply (Glorich Model #WP-AC08030V). The camera is mounted on a Diagnostic Instruments Heavy Duty Boom stand.", "[Camera Setup] At...
91,576
hsqc_metab.nan
5
dx.doi.org/10.17504/protocols.io.5jyl8pd2dg2w/v2
https://www.protocols.io/view/hsqc-metab-nan-c5nyy5fw
NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison
TITLE: hsqc_metab.nan AUTHORS: NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison [DESCRIPTION] This is a protocol for running the Bruker pulse program "hsqcetgpsisp2". [BEFORE_START] This protocol assumes: Your sample is loaded, locked, tuned for ...
["[Create a new dataset]", "[Create a new dataset] A new window opens. On the right top bar, select\nSource = /opt/NAN_METAB/par\n \nIn the list, select the one you want to use:\n\nFor serum and plasma samples:\nHSQC_br600_serum.par: Parameter set using an acquisition mode \"traditional planes\"\nHSQC_NUS_br600_serum.p...
null
null
null
dx.doi.org/10.17504/protocols.io.nxsdfne
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Counting cells with a hemocytometer is an easy way to determine relatively accurate numbers of viable cells.  After determining cell counts, cells can be passaged, frozen away, or used for an experiment at a particular density.  </p> [GUIDELINES] <p>Keep cells sterile by wea...
[]
88,807
UPitt TriState SenNet TMC Cryopreserved Tissue Grinding
1
dx.doi.org/10.17504/protocols.io.q26g7px41gwz/v1
https://www.protocols.io/view/upitt-tristate-sennet-tmc-cryopreserved-tissue-gri-c2yfyftn
Marta Bueno
TITLE: UPitt TriState SenNet TMC Cryopreserved Tissue Grinding AUTHORS: Marta Bueno [DESCRIPTION] This document outlines the tissue pulverization protocol of lung and heart specimens at the TriState SenNet TMC Biospecimen Core at the University of Pittsburgh, as part of the Cellular Senescence Network Program (SenNet)...
["[Tissue pulverization] Pour liquid nitrogen into a mortar and pestle until frozen.", "[Tissue pulverization] Remove tissue from tube and place into liquid nitrogen with a clean cold spatula (precool in liquid\nnitrogen).", "[Tissue pulverization] Grind up the sample with liquid nitrogen using the mortar and pestle.",...
71,463
Co-existing OSA reduce Nuss surgery efficacy
3
dx.doi.org/10.17504/protocols.io.n2bvj86rbgk5/v1
https://www.protocols.io/view/co-existing-osa-reduce-nuss-surgery-efficacy-ch2ft8bn
Mei-Chen Yang
TITLE: Co-existing OSA reduce Nuss surgery efficacy AUTHORS: Mei-Chen Yang [DESCRIPTION] Nuss surgery is effective for pectus excavatum (PE), with a recurrence rate of 1.2–27%. Re-do surgery is effective, but still has a 6% failure rate. Patients with obstructive sleep apnea (OSA) experience repetitive PE-associated s...
[]
70,953
Modified one-step growth (mOSG) assay
4
dx.doi.org/10.17504/protocols.io.4r3l2op9pv1y/v2
https://www.protocols.io/view/modified-one-step-growth-mosg-assay-chiht4b6
Eva JP. Lievens
TITLE: Modified one-step growth (mOSG) assay AUTHORS: Eva JP. Lievens [DESCRIPTION] This protocol describes a modified one-step growth assay, as presented in the manuscript "Life history diversity and signals of trade-offs in a large group of chloroviruses" (Lievens et al., bioRxiv). It is designed to quantify the ads...
["[Preparation (can be done the day beforehand)] Prepare the 1:1000 dilution plates.", "[Preparation (can be done the day beforehand)] Prepare the 1:10000 dilution plates.", "[Preparation (can be done the day beforehand)] If following the alternative 1:1000 dilution option (see step 9.1):\nPrepare the 1:10 intermediate...
18,459
Protocols for activity changes in Neuron-Astrocyte Networks in Culture Under the Effect of Norepinephrine
null
dx.doi.org/10.17504/protocols.io.v93e98n
null
Yasmin Bar El, Sivan Kanner
TITLE: Protocols for activity changes in Neuron-Astrocyte Networks in Culture Under the Effect of Norepinephrine AUTHORS: Yasmin Bar El, Sivan Kanner [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Protocols for Primary cortical neuronal-astrocyte cell culture, Isolated astrocyte cell culture,...
[]
93,561
External Quality Control for inter-Batch comparison (English)
1
dx.doi.org/10.17504/protocols.io.dm6gp36ydvzp/v1
https://www.protocols.io/view/external-quality-control-for-inter-batch-compariso-c7kzzkx6
Ricardo M. Borges
TITLE: External Quality Control for inter-Batch comparison (English) AUTHORS: Ricardo M. Borges [DESCRIPTION] External Quality Control (QCExt): The sample containing the strain code CCMR0280, cultivated in parallel with all batches, will serve as a reference for comparative studies between the strains of the CCMR (Cul...
["[Material] 2 mL microtubes with screw cap\n50 mL Falcon type centrifuge tube\nAnalytical balance (AUW220, Shimadzu)", "[Material in Stock] Initially, a starting batch composed of 25 replicas of the cultivation of the strain code CCMR0280 was (i) cultivated, (ii) collected and combined, (iii) subjected to lyophilizat...
null
null
null
dx.doi.org/10.17504/protocols.io.pnddma6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Rich medium comonly used for Escherichia coli growth and maintenance.</p> [STEPS] ?. ?.
[]
96,434
Exp_01 Taza
0
dx.doi.org/10.17504/protocols.io.6qpvr31povmk/v1
https://www.protocols.io/view/exp-01-taza-daes2bee
Mayleth Medina Cárdenas, Estefanía Sauceda Camacho, Sofía Morga Benítez
TITLE: Exp_01 Taza AUTHORS: Mayleth Medina Cárdenas, Estefanía Sauceda Camacho, Sofía Morga Benítez [DESCRIPTION] Este experimento consiste en la medición de la temperatura de 250 mL de agua contenida en una taza, desde que el agua se encuentra a una temperatura de 84ºC, hasta llegar a una temperatura ambiente de 22ºC...
["[Circuito Arduino y software] Armar el siguiente circuito:", "[Circuito Arduino y software] Conectar el cable USB 2.0 a la placa de Arduino y a la laptop.", "[Experimentación] Hervir agua y con esta vaciar por completo una tasa de 250 mL", "[Resultados de mediciones] Las mediciones registradas en el archivo .txt obte...
81,814
TS Procure 812 - primary fixation Karnovsky&#39;s - transwells (TM - 013)
4
dx.doi.org/10.17504/protocols.io.e6nvwjwpwlmk/v1
https://www.protocols.io/view/ts-procure-812-primary-fixation-karnovsky-39-s-tra-ct5wwq7e
sandra.crameri
TITLE: TS Procure 812 - primary fixation Karnovsky&#39;s - transwells (TM - 013) AUTHORS: sandra.crameri [DESCRIPTION] The method is used to Karnovsky's EM fixed transwells to Procure 812 resin blocks. The presence of 4% paraformaldehyde is only used to satisfy microsecurity requirements. [GUIDELINES] Sample is not o...
["[HEADER] SAN:\n\n\n\n\nSPEC No:\n\n\n\n\nOPERATOR & STEPS:\n\n\n\n\nOPERATOR & STEPS:", "[Conventional] Karnovsky's fixative: 2.5 % volume and 4 % volume in 0.1 Molarity (M) Sorenson's Phosphate buffer (pH 7.2, 300mosmol/kg) for at least 40 min or overnight", "[Conventional] 0.1 Molarity (M) buffer 15 min", "[Co...
99,994
HEPES-Sucrose Cutting Solution
1
dx.doi.org/10.17504/protocols.io.5jyl8peq8g2w/v2
https://www.protocols.io/view/hepes-sucrose-cutting-solution-ddv2268e
Allen Institute
TITLE: HEPES-Sucrose Cutting Solution AUTHORS: Allen Institute [DESCRIPTION] This protocol is used for preparing HEPES-Sucrose Cutting Solution, used for animal perfusion following anesthesia. Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institut...
[]
92,922
Terrific Broth Medium
1
dx.doi.org/10.17504/protocols.io.x54v9pb24g3e/v1
https://www.protocols.io/view/terrific-broth-medium-c6y2zfye
is Sparrow, Steven J Burgess
TITLE: Terrific Broth Medium AUTHORS: is Sparrow, Steven J Burgess [DESCRIPTION] Tartoff, K.D. and Hobbs, C.A. (1987) Improved Media for Growing Plasmid and Cosmid Clones. Bethesda Research Laboratories Focus, 9, 12. [GUIDELINES] Prepare the stock solutions and autoclave separately before combining when cooled to cre...
["[Prepare stock solutions] Combine the following components for 1L TB buffer\n24 g \n20 g \n4 mL Glycerol\nMake up volume to 900 mL with dH2O\n\nAutoclave 121 °C 30 min", "[Prepare medium] Allow solutions to cool to 60 °C then add\n100 mL of 10x TB salts to 900 mL TB buffer.", "[Prepare medium] Store at Room tempe...
null
null
null
dx.doi.org/10.17504/protocols.io.dqh5t5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] For use in "Isolation of cyanophages by plaque assyays" [BEFORE_START] Prepare 100 mL portions of 0.4 to 0.5% (w/v) of purified agar, agarose or low-melting point (LMP) agarose (i.e., Invitrogen #15517-022) in your media of choice. Although LMP agarose can be quite expensive, i...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.fyrbpv6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <div class="page" title="Page 1"> <div class="layoutArea"> <div class="column"> <p>The SensiFAST™ HRM Kit has been developed for fast, highly reproducible High Resolution Melt (HRM) analysis and has been validated on commonly used real-time instruments. A combination of the late...
[]
91,524
Isolation of natural killer (NK) cells from human blood products
1
dx.doi.org/10.17504/protocols.io.81wgbp94yvpk/v3
https://www.protocols.io/view/isolation-of-natural-killer-nk-cells-from-human-bl-c5mcy42w
Philippa R Kennedy
TITLE: Isolation of natural killer (NK) cells from human blood products AUTHORS: Philippa R Kennedy [DESCRIPTION] Standard isolation procedure for peripheral blood mononuclear cells (PBMC) from human blood leukopheresis products. [GUIDELINES] One trimacone will generally provide 500 million - 2 billion PBMCs. NK c...
["[Overview] Healthy donor blood products (Leukopaks) were obtained from Memorial Blood Bank (Minneapolis, MN). All samples were de-identified and their use was approved by the University of Minnesota and NMDP institutional review board in accordance with the Declaration of Helsinki.", "[Overview] Peripheral blood mono...
51,739
Digital Scanning Recommended Practice
1
dx.doi.org/10.17504/protocols.io.bwr3pd8n
https://www.protocols.io/view/digital-scanning-recommended-practice-bwr3pd8n
Patrick Walston, Madalyn Massey
TITLE: Digital Scanning Recommended Practice AUTHORS: Patrick Walston, Madalyn Massey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A protocol regarding digitization of documents stored in paper format. Topics covered: minimum standards, best practices, digital scanning hardware, software, file st...
["[Introduction]\nWhat is it?Digitization is the process of turning physical information into digital data. The USGS and National Geological and Geophysical Data Preservation Program (NGGDPP), aim to preserve and curate historical geological information using Digitization. By following standard processes, scientists ca...
34,196
Protein expression in Bacillus subtilis
1
dx.doi.org/10.17504/protocols.io.bdmui46w
https://www.protocols.io/view/protein-expression-in-bacillus-subtilis-bdmui46w
Kristoffer Bach Falkenberg, Cristina Hernandez Rollan, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm
TITLE: Protein expression in Bacillus subtilis AUTHORS: Kristoffer Bach Falkenberg, Cristina Hernandez Rollan, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-style:italic;">B. subtilis </span><span>is a gram-positive bacteria u...
["[Cal18-2 media preparations]\nPrepare a stock solution of 2.0g/L Na2MoO4. Sterilize by filtration", "[Cal18-2 media preparations]\nAdd a magnetic stirrer to the blue cap bottle and place the bottle on a stirring plate. Turn on the stirring, and make sure it's mixing well.", "[Cal18-2 media preparations]\nAdd the foll...
67,518
A versatile nuclei extraction protocol for single nucleus sequencing in non model species – optimization in various Atlantic salmon tissues.
4
null
https://www.protocols.io/view/a-versatile-nuclei-extraction-protocol-for-single-cd66s9he
Rose Ruiz Daniels, Richard S Taylor, Ross Dobie, Sarah Salisbury, Emily Clark, Dan Macqueen, Diego Robledo
TITLE: A versatile nuclei extraction protocol for single nucleus sequencing in non model species – optimization in various Atlantic salmon tissues. AUTHORS: Rose Ruiz Daniels, Richard S Taylor, Ross Dobie, Sarah Salisbury, Emily Clark, Dan Macqueen, Diego Robledo [DESCRIPTION] Single cell RNA sequencing has rapidly be...
["[Nucleus isolation workflow for ST-based buffers] on ice, place a piece of frozen tissue into one well of a 6-well tissue culture plate with 1 mL TST.", "[Nucleus isolation workflow for ST-based buffers] on ice, mince tissue initially using Tungsten Carbide scissors for 30 s and then with Noyes Spring Scissors for ...
28,111
Co-Incubation protocol for transforming heterotrophic dinoflagellates (e.g. Oxyrrhis marina)
null
dx.doi.org/10.17504/protocols.io.7pphmmn
null
Lu Wang, Brittany Sprecher, Huan Zhang, and Senjie Lin
TITLE: Co-Incubation protocol for transforming heterotrophic dinoflagellates (e.g. Oxyrrhis marina) AUTHORS: Lu Wang, Brittany Sprecher, Huan Zhang, and Senjie Lin [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Prepare vector and transform into Escherichia coli] Prepare a plasmid carrying proper promo...
["[Prepare vector and transform into Escherichia coli]\nPrepare a plasmid carrying proper promoter, selection marker, and reporter gene. Transform this plasmid into E. coli cells using standard heat shock protocol.", "[Grow transformed E. coli]\nGrow E. coli cells at a volume equal to 1/10 the desired volume of dinofla...
18,638
GYROS
null
dx.doi.org/10.17504/protocols.io.wfnfbme
null
Anish Chhetri
TITLE: GYROS AUTHORS: Anish Chhetri [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">I would absolutley go for StrainControl Laboratory Manager. I find this software extremely good and easy to use. Lets you manage strains, plasmids, oligos, antibodies and inventories.</div></div> [STEPS] ?. [1] Prim...
["[1]\nPrime the instrument 200%", "[1]\nDownload the sequence file", "[1]\nMRD of samples 1:2", "[1]\nCapture Prep\n10 Mass/Volume Percent", "[1]\nDetector Prep", "[1]\nOpen the Gyros Software"]
null
null
null
dx.doi.org/10.17504/protocols.io.m9vc966
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>Background</strong></p> <p>The use of mobile technology such as phone applications (apps) has been proposed as an efficient means of providing health and clinical information in a variety of healthcare settings. We developed the Health-e Babies app as an Android smart...
[]
44,047
BSCI:414--Lab 9: Cloning the SARS-CoV-2 Spike Protein into a E. coli Protein Expression Vector
1
null
https://www.protocols.io/view/bsci-414-lab-9-cloning-the-sars-cov-2-spike-protei-bn9pmh5n
Harley King
TITLE: BSCI:414--Lab 9: Cloning the SARS-CoV-2 Spike Protein into a E. coli Protein Expression Vector AUTHORS: Harley King [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The SARS-CoV-2 spike protein does not express solubly in E. coli cells. This may be due to E. coli's inability to glycosylate...
["[Explore the Spike Protein]\nExplore the structure of the SARS-CoV-2 spike protein using Cov3D.", "[Explore the Spike Protein]\nRead abstract of Zuo et al. concerning previous attempts at expressing SARS-CoV spike protein fused with SUMO tag.", "[Clone spike -sumo into pET28.]\nUsing a Gibson cloning strategy in Benc...
35,043
Generating cloned sheep embryos by zona-free somatic cell transfer
1
dx.doi.org/10.17504/protocols.io.begbjbsn
https://www.protocols.io/view/generating-cloned-sheep-embryos-by-zona-free-somat-begbjbsn
Zachariah Mclean, Sarah Appleby, Lisanne Fermin, Bjorn Oback
TITLE: Generating cloned sheep embryos by zona-free somatic cell transfer AUTHORS: Zachariah Mclean, Sarah Appleby, Lisanne Fermin, Bjorn Oback [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol summarises zona-free sheep cloning by somatic cell transfer (SCT), adapted from a simila...
["[Induce donor cell quiescence by serum starvation]\nInduce donor cells into quiescence (or G0) by serum starvation", "[In vitro maturation of oocytes]\nRetrieve oocytes from slaughterhouse ovaries\nSheep (ovine) is a seasonally reproductive species and SCT experiments are best carried out during the extended breeding...
95,131
BAF_Protocol_005 Database Search Proteome Discoverer into Scaffold
1
dx.doi.org/10.17504/protocols.io.q26g7p28kgwz/v1
https://www.protocols.io/view/baf-protocol-005-database-search-proteome-discover-c853zy8n
nesf
TITLE: BAF_Protocol_005 Database Search Proteome Discoverer into Scaffold AUTHORS: nesf [DESCRIPTION] This protocol lays out the basic steps for taking a Thermo RAW file and doing a standard database search in Proteome Discoverer 2.5+ and putting the search results into Scaffold 5.3+ for display. Included is also the ...
["[Proteome Discoverer 2.5 Database Searching] Thermo RAW files are set up to search in PD 2.5 software (Proteome Discoverer) to produce an output MSF file. The RAW files for an individual project are placed in a folder and a sub folder is created with the MSF files produced by PD 2.5 inside. The MSF files will be load...
null
null
null
dx.doi.org/10.17504/protocols.io.ucwesxe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol is used to dissociate adult (8-10 wk) mouse ear "on ice" (in order to reduce gene expression artifacts). The cell yield is 9000 cells/mg (213000 cells total from 23 mg tissue), with 98% viable. The protocol involves a 2-layered dissociation, incubating on ice with ...
["[Isolating tissue] Dissect out ear tissue and place in ice-cold hypothermosol on ice.", "[Isolating tissue] {\"blocks\":[{\"key\":\"jm3a\",\"text\":\"Using forceps, transfer ear tissue to petri dish on ice. Mince ear tissue thoroughly on ice for 3-4 min on ice into 1-mm3 pieces using razor blade while manipulating ti...
31,737
Methods and protocols from Chaiyarat et al. (2019) for systematic reintroduction of bateng (Bos javanicus)
null
dx.doi.org/10.17504/protocols.io.ba8zihx6
null
Rattanawat Chaiyarat, Namphung Youngpoy, Praeploy Kongsurakan, Seree Nakbun
TITLE: Methods and protocols from Chaiyarat et al. (2019) for systematic reintroduction of bateng (Bos javanicus) AUTHORS: Rattanawat Chaiyarat, Namphung Youngpoy, Praeploy Kongsurakan, Seree Nakbun [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Banteng (</span><span style = "font-style:itali...
[]
56,827
Miraprep: Fast Plasmid Prep
1
dx.doi.org/10.17504/protocols.io.b3q3qmyn
https://www.protocols.io/view/miraprep-fast-plasmid-prep-b3q3qmyn
Zaki Molvi
TITLE: Miraprep: Fast Plasmid Prep AUTHORS: Zaki Molvi [DESCRIPTION] An adaptation of the original miraprep protocol published by Pronobis et al. PLoS One (2016). [STEPS] SECTION: Introduction 1. My adaptation of the miraprep method originally published by Pronobis et al. The miraprep method is highly preferred sinc...
["[Introduction] My adaptation of the miraprep method originally published by Pronobis et al. The miraprep method is highly preferred since plasmid prep kits larger than minipreps take too long and require scarier, louder centrifuges.", "[Protocol] Inoculate an overnight culture in 2xYT and appropriate antibiotic1\nThe...
104,401
Transfections for gene delivery and genome editing in S. rosetta (VERSION 4)
1
dx.doi.org/10.17504/protocols.io.j8nlk86o5l5r/v1
https://www.protocols.io/view/transfections-for-gene-delivery-and-genome-editing-dh7r39m6
David Booth
TITLE: Transfections for gene delivery and genome editing in S. rosetta (VERSION 4) AUTHORS: David Booth [DESCRIPTION] This protocol details the preparation and execution of CRISPR/Cas9 genome editing in S. rosetta. The protocol builds on a method to transfect macromolecules into S. rosetta for delivering a purified ...
["[Culture Cells] Seed a large culture of S. rosetta.", "[Culture Cells] Two days prior to transfection, inoculate 80 mL of media (15% Red Algae + 2% Peptone-Yeast-Glycerol) with a culture of S. rosetta feeding on E. pacifica (ATCC PRA-390) to a final concentration of S. rosetta of 1.5*10^4 cells/ml. Also add 400 µLo...
95,586
Protocol for &#34;ventriculography-assisted stereotaxic surgery in non-human primates
0
dx.doi.org/10.17504/protocols.io.e6nvwd3w7lmk/v1
https://www.protocols.io/view/protocol-for-34-ventriculography-assisted-stereota-c9kaz4se
jlanciego
TITLE: Protocol for &#34;ventriculography-assisted stereotaxic surgery in non-human primates AUTHORS: jlanciego [DESCRIPTION] An stept-by-step procedure describing ventriculography-assisted stereotaxic surgery in non-human primates is provided here, including (i) animal pre-surgical preparation, (ii) stereotaxic surge...
["[Animal preparation] Pre-surgical anesthesia: to be induced with ketamine (5 mg/Kg) and midazolam (0.5 mg/Kg), administered intramuscularly.", "[Animal preparation] Surgical shaving: comprising left and right superior and inferior limbs and thorax for the placement of adhesive EKG electrodes.", "[Animal preparation] ...
52,287
cDNA Synthesis Using SuperScript III First-Strand Synthesis System for RT-PCR
4
dx.doi.org/10.17504/protocols.io.bxa7pihn
https://www.protocols.io/view/cdna-synthesis-using-superscript-iii-first-strand-bxa7pihn
Lynn Doran
TITLE: cDNA Synthesis Using SuperScript III First-Strand Synthesis System for RT-PCR AUTHORS: Lynn Doran [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">The SuperScript® III First-Strand Synthesis System for RT-PCR is used to synthesize first-strand cDNA from purified total RNA. RNA targets from 1...
["Allow reagents to thaw completely, mix, and briefly minicentrifuge 10 mM dNTP mix and 50 ng/ul random hexamers before use. Store on ice when not in use.", "Label two PCR tubes per sample, RT and NRT.\nBest practice for qPCR is to prepare a no reverse transcriptase (NRT) reaction for each of your samples to ensure tha...
null
null
null
dx.doi.org/10.17504/protocols.io.c2dya5
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] This protocol is designed to purify single- or double-stranded DNA fragments from PCR and other enzymatic reactions (see page 8). For cleanup of other enzymatic reactions, follow the protocol as described for PCR samples or use the MinElute Reaction Cleanup Kit. Fragments rangin...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.mayc2fw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><strong>A protocol for isolation of RNA from blood and RT-PCR for detection of live Brucella organisms in cattle</strong></p> [STEPS]
[]
70,086
LRRK2 Staining in mouse brain sections 
4
dx.doi.org/10.17504/protocols.io.e6nvwj64dlmk/v1
https://www.protocols.io/view/lrrk2-staining-in-mouse-brain-sections-cgpetvje
Chuyu Chen
TITLE: LRRK2 Staining in mouse brain sections  AUTHORS: Chuyu Chen [DESCRIPTION] LRRK2 is a widely expressed, multifunctional protein.LRRK2 mutations are the major cause to inherited and sporadic PD. Staining of LRRK2 in mouse brain can be challenging. This protocol provides an optimised way to achieve LRRK2 staining ...
["Prepare brain cryosections in 20μm thickness", "Blocking with 5% serum (preferably same serum as host of secondary antibody) 30 min RT", "Wash with PBS 5 min 2x", "Permeabilize with 0.5% Triton 15 min RT", "Wash with PBS 5 min 2x", "VECTOR® antigen unmasking solution 48ul/5ml water (in small glass sample vial). Micr...
15,196
K and K+Si medium
null
dx.doi.org/10.17504/protocols.io.s34egqw
null
Roscoff Culture Collection
TITLE: K and K+Si medium AUTHORS: Roscoff Culture Collection [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Medium to grow most phytoplankton species in small eukaryotes (Mamiellophyceae, Pelagophyceae etc...).</div><div class = "text-block">When using for diatoms and coccolithophorids add silica (...
["Filter 1L of old seawater of at least two months on prefilter and filter 0,2 microns", "Heat seawater during 20min at 100°C", "Under hood, to seawater, add these nutriments that have been autoclaved (excepted vitamin) : ABC1Quantity\nCompound\nStock Solution\n21.0 mL\nSodium Nitrate (NaNO3)\n75.0 g/L of H2O\n31.0 mL...
null
null
null
dx.doi.org/10.17504/protocols.io.n43dgyn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>A real-time PCR targeting the DNA-dependent RNA polymerase of Orthopoxviruses.</p> <p>This protocol was designed and developed at this laboratory.</p> [GUIDELINES] <ul> <li>If using a different brand or model of real-time thermocycler, check the concentration of ROX is adequ...
[]
88,426
JMN-MSMP Muscle Bulk RNAseq
1
dx.doi.org/10.17504/protocols.io.rm7vzxjz4gx1/v2
https://www.protocols.io/view/jmn-msmp-muscle-bulk-rnaseq-c2kiycue
ccherry
TITLE: JMN-MSMP Muscle Bulk RNAseq AUTHORS: ccherry [DESCRIPTION] RNA sequencing of muscle [STEPS] SECTION: Bulk RNA sequencing 1. cDNA synthesis and library preparation were performed using the TruSeq RNA Library Prep Kit V2 following the manufacturer's protocol. SECTION: Bulk RNA sequencing 2. The Agilent BioAnaly...
["[Bulk RNA sequencing] cDNA synthesis and library preparation were performed using the TruSeq RNA Library Prep Kit V2 following the manufacturer's protocol.", "[Bulk RNA sequencing] The Agilent BioAnalyzer system was used to perform quality control on samples during the library prep process as suggested in the manufac...
48,205
IHC AD neuropathology protocol 
4
dx.doi.org/10.17504/protocols.io.btbmnik6
https://www.protocols.io/view/ihc-ad-neuropathology-protocol-btbmnik6
Christiana Bjorkli
TITLE: IHC AD neuropathology protocol  AUTHORS: Christiana Bjorkli [DESCRIPTION] Immunohistochemical processing was conducted on tissue from chronically implanted animals, as well as non-implanted transgenic mice for characterization of AD neuropathology. [STEPS] SECTION: Day 1 fluorescent IHC 1. Heat-induced antigen...
["[Day 1 fluorescent IHC] Heat-induced antigen retrieval on all tissue at 60 °C for 2 hours in phosphate buffer (PB).", "[Day 1 fluorescent IHC] Wash sections 3 x 10min in PB containing 0.2 % Triton X-100 (PBT+).", "[Day 1 fluorescent IHC] Block sections using 5 % normal goat serum in PBT+ for 1 hour.", "[Day 1 fluor...
54,354
Revival of S. elongatus from plates
4
dx.doi.org/10.17504/protocols.io.bzbsp2ne
https://www.protocols.io/view/revival-of-s-elongatus-from-plates-bzbsp2ne
Akashdutta
TITLE: Revival of S. elongatus from plates AUTHORS: Akashdutta [DESCRIPTION] On receiving wild type or tranformed S. elongatus plates, one needs to revive them for further studies. Revival of S. elongatus involves making its liquid culture in BG-11 medium. The following protocol describes the liquid culture preparat...
["Take a 50 mL autoclaved flask.", "Take a 10 mL aliquot of BG-11 solution in a falcon tube and transfer it into the flask. Keep the flask aside", "Dip the pipette tip in the aliquot of the BG-11 medium and gently mix.", "Take the cyanobacteria plate and gently scrape it with the tip of a 1 mL loaded micropipette.", ...
57,027
Acetylation of lysines on affinity-purification matrices to reduce co-digestion of bead-bound ligands
4
dx.doi.org/10.17504/protocols.io.kxygxzexkv8j/v2
https://www.protocols.io/view/acetylation-of-lysines-on-affinity-purification-ma-b3xbqpin
David M. Hollenstein, Margarita Maurer-Granofszky, Dorothea Anrather, Thomas Gossenreiter, Natascha Hartl, Markus Hartl
TITLE: Acetylation of lysines on affinity-purification matrices to reduce co-digestion of bead-bound ligands AUTHORS: David M. Hollenstein, Margarita Maurer-Granofszky, Dorothea Anrather, Thomas Gossenreiter, Natascha Hartl, Markus Hartl [DESCRIPTION] In mass-spectrometry-based interaction proteomics on-bead digestion...
["[Acetylation protocol] Wash beads 3x with 100µl Reaction buffer, using the magnetic rack. \nRemove supernatant after the final wash.", "[Acetylation protocol] Add 19 µl Reaction buffer and 1 µl Sulfo-NHS-Acetate (100 mM) to obtain a final concentration of 5 mM Sulfo-NHS-Acetate. \nIncubate 1h at room temperature with...
52,480
Microfluidics Lithography 1 Mold Fabrication: Spin Coating of Photoresist (version s2)
1
dx.doi.org/10.17504/protocols.io.bxg8pjzw
https://www.protocols.io/view/microfluidics-lithography-1-mold-fabrication-spin-bxg8pjzw
C. Yunus Sahan, Serhat S
TITLE: Microfluidics Lithography 1 Mold Fabrication: Spin Coating of Photoresist (version s2) AUTHORS: C. Yunus Sahan, Serhat S [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:center"></div></div><div style = "text-align :; float : ;"><img style = "" src =...
["[SpinCoater Instrument Adjustment]\nDispense 1ml of photoresist (SU8) resin for each inch (25mm) of Si wafer substrate diameter.Spin at 500 rpm for 10 seconds with an acceleration of 100 rpm/second.Spin at \"N\" rpm for 30 seconds with an acceleration of 300 rpm/second.Expected results of photoresist thickness: 40u...
null
null
null
dx.doi.org/10.17504/protocols.io.khxct7n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Adapted from Core LJ, Waterfall JJ, Lis JT. Nascent RNA sequencing reveals widespread pausing and divergent initiation at human promoters. Science. 2008;322(5909):1845-8. PubMed PMID: 19056941.</p> <p>This method can be used to detect nascent transcripts. The conditions given...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.cpkvkv
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
107,703
Tissue preparation and tissue imaging for MERFISH
0
dx.doi.org/10.17504/protocols.io.14egn6opyl5d/v1
https://www.protocols.io/view/tissue-preparation-and-tissue-imaging-for-merfish-dmex43fn
Cameron Oram
TITLE: Tissue preparation and tissue imaging for MERFISH AUTHORS: Cameron Oram [DESCRIPTION] MERSCOPE Sample preparation protocol and imaging protocol [STEPS] SECTION: Perfusion and sectioning of brain 1. C57Bl6 male mice were housed in groups until 2-3 months of age. SECTION: Perfusion and sectioning of brain 2. 4% ...
["[Perfusion and sectioning of brain] C57Bl6 male mice were housed in groups until 2-3 months of age.", "[Perfusion and sectioning of brain] 4% PFA solution was made from 32% PFA concentrate in 1x PBS.", "[Perfusion and sectioning of brain] Mice were perfused first with 25 mL of chilled 1x PBS (DNAse, RNAse free) follo...
32,625
Surgical Isolation of Stellate Ganglia and Electrophysiology
1
dx.doi.org/10.17504/protocols.io.bb4riqv6
https://www.protocols.io/view/surgical-isolation-of-stellate-ganglia-and-electro-bb4riqv6
Mark Doyle
TITLE: Surgical Isolation of Stellate Ganglia and Electrophysiology AUTHORS: Mark Doyle [DESCRIPTION] Surgical Isolation of Stellate Ganglia and Electrophysiology [STEPS] 1. The thorax of adult mice of either sex were removed under deep Isoforane anesthesia according to approved protocols by the Institutional Animal...
["The thorax of adult mice of either sex were removed under deep Isoforane anesthesia according to approved protocols by the Institutional Animal Care and Use Committee at Oregon Health and Science University, and guidelines of the National Institutes of Health. The heart and SG were perfused with modified Ringers that...
41,462
R editing of HMMSEARCH output: heatmap with associated with gene tree and protein illustration
5
dx.doi.org/10.17504/protocols.io.bkqwkvxe
https://www.protocols.io/view/r-editing-of-hmmsearch-output-heatmap-with-associa-bkqwkvxe
Kanae Nishii
TITLE: R editing of HMMSEARCH output: heatmap with associated with gene tree and protein illustration AUTHORS: Kanae Nishii [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">R codes used for phyml tree and profile HMM heatmap visualization, protein domain illustration</div></div> [STEPS] ?. [R librar...
["[R libraries]\n#libraries for tree drawinglibrary(ggtree)library(ape)library(phytools)#libraries for heatmaplibrary(reshape2)library(dplyr)library(ggplot2)library(aplot)", "[tree drawing]\n#rooting tree#midpoint rooted treetree2 #rooted by outgroup samplestree2 #check tree2ggtree(tree2)", "[tree drawing]\n#read Newic...
91,658
Visium Direct Mount Fresh Frozen -- University of Minnesota TMCs
1
dx.doi.org/10.17504/protocols.io.6qpvr3qdovmk/v1
https://www.protocols.io/view/visium-direct-mount-fresh-frozen-university-of-min-c5riy54e
Laura J Niedernhofer, David A Bernlohr
TITLE: Visium Direct Mount Fresh Frozen -- University of Minnesota TMCs AUTHORS: Laura J Niedernhofer, David A Bernlohr [DESCRIPTION] The Visium Spatial Gene Expression Solution measures the total mRNA in tissue sections and requires a Visium Spatial slide with intact tissue sections as input. This protocol outlines m...
["[Tissue Optimization]", "[Fixation, H&amp;E Staining &amp; Imaging]", "[Library Preparation &amp; Sequencing]", "[FASTQ Generation] BCL data from Illumina sequencer is demultiplexed and converted into FASTQ format using bcl2fastq version 2.20.0"]
92,592
Populating NCBI template for submissions using BioNumerics
1
dx.doi.org/10.17504/protocols.io.36wgq3ejylk5/v2
https://www.protocols.io/view/populating-ncbi-template-for-submissions-using-bio-c6nqzddw
Ruth Timme, Maria Balkey, Julie Haendiges, Brian Sauders, Tina.Pfefer
TITLE: Populating NCBI template for submissions using BioNumerics AUTHORS: Ruth Timme, Maria Balkey, Julie Haendiges, Brian Sauders, Tina.Pfefer [DESCRIPTION] PURPOSE: to define the standard operating procedure for collecting isolate metadata using BioNumerics for submission of food/environmental isolates to NCBI. ...
["Metadata SampleSheet preparation \n\nBefore uploading your sequencing run or linking NCBI sequencing records at the BioNumerics platform make sure to fill out the metadata spreadsheet form. \n\nPlease download the template and guidelines included in the file \n\nCreate the fields NCBI_bioproject, Attribute_package...
44,592
Functional Analysis of Membrane Proteins Produced by Cell-Free Translation
4
dx.doi.org/10.17504/protocols.io.bpsqmndw
https://www.protocols.io/view/functional-analysis-of-membrane-proteins-produced-bpsqmndw
Srujan Kumar Dondapati, Doreen A. Wüstenhagen, Stefan Kubick
TITLE: Functional Analysis of Membrane Proteins Produced by Cell-Free Translation AUTHORS: Srujan Kumar Dondapati, Doreen A. Wüstenhagen, Stefan Kubick [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Cell-free production is a valuable and alternative method for the synthesis of membrane protei...
["[3.1 Preparation of Nanodiscs (NDs)]\nFirst prepare () containing , with and .\n[MSP protein solution]\n[Tris]\n[NaCl]\n[EDTA]", "[3.1 Preparation of Nanodiscs (NDs)]\nDissolve DMPC, DOTAP and DOPG lipids in cholate buffer at a concentration of (see Note 2).", "[3.1 Preparation of Nanodiscs (NDs)]\nSet up the rea...
40,387
Effective identification of RNA-binding proteins using RNA Immunoprecipitation
4
dx.doi.org/10.17504/protocols.io.bjpbkmin
https://www.protocols.io/view/effective-identification-of-rna-binding-proteins-u-bjpbkmin
Georgios I Laliotis, Philip N. Tsichlis
TITLE: Effective identification of RNA-binding proteins using RNA Immunoprecipitation AUTHORS: Georgios I Laliotis, Philip N. Tsichlis [STEPS] ?. [Chemicals Required] For 100ml1.28M sucrose 43.21g .justify:after { content: ""; display:inline-block; width: 100%; ...
["[Chemicals Required]\nFor 100ml1.28M sucrose 43.21g\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\t1M Tris-HCL 7.5 4ml\n\t\t\t\t\t\t\t .justify:after {\n\t\t\t\t\t\t\t cont...
50,707
Intrahepatic implantation of tumor cells
4
dx.doi.org/10.17504/protocols.io.bvrtn56n
https://www.protocols.io/view/intrahepatic-implantation-of-tumor-cells-bvrtn56n
Shubhangi Agarwal, Robert Bok, donna.peehl , Renuka Sriram
TITLE: Intrahepatic implantation of tumor cells AUTHORS: Shubhangi Agarwal, Robert Bok, donna.peehl , Renuka Sriram [DESCRIPTION] This protocol describes the steps required for successful implantation of small cell neuroendocrine prostate cancer patient-derived xenograft (PDX) cells in the liver. Liver is one of...
["[Preparation before surgery] Preparation of surgical instruments and supplies: All of the instruments and supplies should be sterilized.", "Preparation of the PDX cells for implantation", "[Preparation of cells for implantation from frozen biobank] Retrieve cryovials containing cells from liquid nitrogen storage. Tha...
45,459
m9_media
4
null
https://www.protocols.io/view/m9-media-bqmtmu6n
Elizabeth Fozo
TITLE: m9_media AUTHORS: Elizabeth Fozo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">M9 media protocols</div></div> [STEPS] ?. [M9 minimal media: low osmolarity media for E. coli, resulting in slower growth rate] Mix the ingredients (forms white precipitate but is dissolved after steering), filt...
["[M9 minimal media: low osmolarity media for E. coli, resulting in slower growth rate]\nMix the ingredients (forms white precipitate but is dissolved after steering), filter sterilize, and store at room temperature.M9 Salts1X5X200100Magnesium sulfate2mM1M21Calcium chloride0.1mM1M0.10.05Vitamin B11ug/mL10mg.mL0.10.05Ca...
76,661
Illumina Amplicon Sequencing using two step PCR
1
null
https://www.protocols.io/view/illumina-amplicon-sequencing-using-two-step-pcr-cn4vvgw6
Lindsay Kate Newbold, Joetay, Jonathan Warren, Daniel S Read, kerry.walsh, Amy Thorpe
TITLE: Illumina Amplicon Sequencing using two step PCR AUTHORS: Lindsay Kate Newbold, Joetay, Jonathan Warren, Daniel S Read, kerry.walsh, Amy Thorpe [DESCRIPTION] This document is designed to provide the researcher with all of the information required to undertake two step amplicon sequencing using the Illumina MiSeq...
["[Nextera index plates- Prepared prior to Step 1 PCR] Order indexing primers direct from oligo manufacturer (For example IDT, Sigma genosys or MWG) suspended in water 0.5 µM scale, Desalt purification and10 µM concentration. These indexing primers consist of: an Illumina Nextra adapter i5 (Forward primer) AATGATACGG...
91,221
Housing and Care for Hymenochirus boettgeri
1
dx.doi.org/10.17504/protocols.io.ewov1qxdpgr2/v1
https://www.protocols.io/view/housing-and-care-for-hymenochirus-boettgeri-c5bvy2n6
Tamilie Carvalho, Catherine Si, Rebecca A. Clemons, Evelyn Faust, Timothy Y. James
TITLE: Housing and Care for Hymenochirus boettgeri AUTHORS: Tamilie Carvalho, Catherine Si, Rebecca A. Clemons, Evelyn Faust, Timothy Y. James [DESCRIPTION] This protocol provides an overview of housing and care procedures for H. boettgeri in a controlled laboratory environment, encompassing feeding at all life stages...
["[Housing] Juvenile or adult frogs", "[Housing] Animals can be kept in groups or individually as needed. House animal groups in 38 L glass tanks (standard system; Figure 1A) with a density of at least 1.5 L of water per individual. House animals individually in 1 L plastic tanks (static system; Figure 1B).", "[Housing...
null
null
null
dx.doi.org/10.17504/protocols.io.gdfbs3n
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Find the words in common between two files.  It is suggested to use hashes, but Sets and Bags would also useful for the more advanced student.</p> [STEPS] ?. ?. ?.
[]
29,050
Folding of FluoroCubes
1
dx.doi.org/10.17504/protocols.io.8k2huye
https://www.protocols.io/view/folding-of-fluorocubes-8k2huye
Stefan Niekamp, Nico Stuurman, Ronald D. Vale
TITLE: Folding of FluoroCubes AUTHORS: Stefan Niekamp, Nico Stuurman, Ronald D. Vale [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Photobleaching limits extended imaging of fluorescent biological samples. Here, we developed DNA based “FluoroCubes” that are similar in size to the green fluorescent ...
["[Design]\nPick a dye that you would like to use for the six dye FluoroCube. We have tested ATTO 488, ATTO 565, ATTO 647N, Cy3, Cy3N (sulfonated Cy3), and Cy5. Based on our experience, ATTO 647N and Cy3N work very well as six dye FluoroCubes and we recommend using these two dyes.", "[Design]\nOrder the modified oligon...
43,539
Tetraspanin (CD9, CD63, CD81) Western Blot for Extracellular Vesicles
4
dx.doi.org/10.17504/protocols.io.bnrtmd6n
https://www.protocols.io/view/tetraspanin-cd9-cd63-cd81-western-blot-for-extrace-bnrtmd6n
Dima Ter-Ovanesyan, Wendy Trieu , Maia Norman, Roey Lazarovits, George Church, David Walt
TITLE: Tetraspanin (CD9, CD63, CD81) Western Blot for Extracellular Vesicles AUTHORS: Dima Ter-Ovanesyan, Wendy Trieu , Maia Norman, Roey Lazarovits, George Church, David Walt [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Extracellular vesicles (EVs) are released by all mammalian cells and are tho...
["[Day 1 ]\nAdd Bolt 4x LDS sample buffer to samples for a final concentration of 1x LDS or resuspend in 1x LDS directly if the samples are pellets.", "[Day 1 ]\nVortex, spin down, and denature the samples in heat block at 70 °C for 10 minutes.", "[Day 1 ]\nFill the Western Blot mini gel tank with 1x MES running buffer...
39,175
Statistical Analysis Plan for Development of Injury Prediction Models for the MP3 Return to Duty Study
1
dx.doi.org/10.17504/protocols.io.bihfkb3n
https://www.protocols.io/view/statistical-analysis-plan-for-development-of-injur-bihfkb3n
Dan Rhon, Tina Greenlee, Jeff Sorensen, Garrett Bullock
TITLE: Statistical Analysis Plan for Development of Injury Prediction Models for the MP3 Return to Duty Study AUTHORS: Dan Rhon, Tina Greenlee, Jeff Sorensen, Garrett Bullock [STEPS] ?. [Adminstrative Information] ADMINISTRATIVE INFORMATIONThe SAP below is written following the Guidelines for Content of Statistical An...
["[Adminstrative Information]\nADMINISTRATIVE INFORMATIONThe SAP below is written following the Guidelines for Content of Statistical Analysis Plans in Clinical Trials (adapted for the purposes of this cohort study) JAMA Reference: http://dx.doi.org/10.1001/jama.2017.18556", "[Adminstrative Information]\nTitle and Tria...