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35,160
Multi-Seq: my notes from the lab
null
dx.doi.org/10.17504/protocols.io.bejyjcpw
https://www.protocols.io/view/multi-seq-my-notes-from-the-lab-bejyjcpw
Luciano Martelotto
TITLE: Multi-Seq: my notes from the lab AUTHORS: Luciano Martelotto [DESCRIPTION] <div class = "text-blocks"><table border><tr style = "text-align:center;"><td> </td><td>A</td></tr><tr><td style = "text-align:center;">1</td><td rowspan = "" colspan = "" style ="display : table-cell;">Please note these are just my not...
["First thing to do is to get very clean nuclei preps. You may start with less clean nuclei if you like but it is likely that CMO will bind to the debris, and that is not good (just a thought). So, if you work with cell lines, iPSCs, PBMCs or some of the ‘liquid’ cancers then nuclei will be pretty clean, but depending ...
null
null
null
dx.doi.org/10.17504/protocols.io.tqdems6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Immunocytochemistry for morphology analysis. Staining of cell culture in order to obtain fluorescence images of the culture. [STEPS] SECTION: Cell fix ?. SECTION: Staining ?. SECTION: Permeabilization ?. SECTION: Block ?. SECTION: Primary antibody incubation ?. SECTION: Se...
["[Cell fix] {\"blocks\":[{\"key\":\"bq65e\",\"text\":\"1. Wash the culture twice in a phosphate buffered saline (PBS);\",\"type\":\"unstyled\",\"depth\":0,\"inlineStyleRanges\":[],\"entityRanges\":[],\"data\":[]},{\"key\":\"6g3ps\",\"text\":\"2. Fix with 4% paraformaldehyde (Merck) for 10 min , then put it in PBS unti...
5,602
Transformation of E.coli with pGem-T Easy
null
dx.doi.org/10.17504/protocols.io.hqab5se
null
Gustaf Degen
TITLE: Transformation of E.coli with pGem-T Easy AUTHORS: Gustaf Degen [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [A-tailing procedure (Day 1)] Because the Phusion polymerase will remove the A-overhangs, adding these with Taq DNA Polymerase is necessary, as this is required for cloningMix the followi...
["[A-tailing procedure (Day 1)]\nBecause the Phusion polymerase will remove the A-overhangs, adding these with \nTaq DNA Polymerase is necessary, as this is required for cloningMix the following reagents: AB1PCR Product1-7µl210x Taq Buffer1µl3ATP to 0.2mM1µl4Taq DNA Pol 5U1µl5dH2Oto 10µlIncubate for 30 min at 70°CIdea...
null
null
null
dx.doi.org/10.17504/protocols.io.nwmdfc6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Testing antibacterial activity of bacteriophages on hard surfaces</p> [BEFORE_START] <p> </p> <p>The following materials are needed:</p> <p>- individual pure bacterial cultures</p> <p>- bacteriophage preparation (either commercial or home-prepared)</p> <p>- bacterial growth ...
[]
94,282
α-Synuclein sedimentation assay
4
dx.doi.org/10.17504/protocols.io.6qpvr3nwpvmk/v1
https://www.protocols.io/view/synuclein-sedimentation-assay-c8bizske
arpine.sokratian
TITLE: α-Synuclein sedimentation assay AUTHORS: arpine.sokratian [DESCRIPTION] The protocol describes step-by-step instructions on how to set a sedimentation assay to monitor the aggregation of alpha-synuclein over time. The protocol allows for precise quantification of fibril formation kinetics using SDS-PAGE. This ...
["[Amplification set-up] Thaw down a-syn monomer and/or sonicated fibril aliquots. Use water-bath.\nDo not generate bubbles by pipetting or shaking", "[Amplification set-up] Measure monomer concentration via Nanodrop\nAdd Amount of 5x, 10x or 20x diluted aliquots in PBS onto nanodrop piedestal; \nParameters: other pro...
null
null
null
dx.doi.org/10.17504/protocols.io.dzs76d
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] The following procedure is an adaptation of the original FLVP protocol from Hennes et al. (1995) for use with SYBR Green I stain. Normally, this protocol will take 2 days given that the softening of the viral pellet is an overnight step. However, if the viral pellet redissolves ...
[]
62,255
Keto Start ACV Gummies - Most Effective Weight Loss Gummies - Best Gummies For Weight Loss?( 2022 Update)
3
dx.doi.org/10.17504/protocols.io.q26g74drqgwz/v1
https://www.protocols.io/view/keto-start-acv-gummies-most-effective-weight-loss-b82prydn
Keto Start ACV Gummies
TITLE: Keto Start ACV Gummies - Most Effective Weight Loss Gummies - Best Gummies For Weight Loss?( 2022 Update) AUTHORS: Keto Start ACV Gummies [DESCRIPTION] Keto Start ACV Gummies [STEPS]
[]
53,826
Crosslinking assay to study a specific cargo-coat interaction through a transmembrane receptor in the secretory pathway
4
dx.doi.org/10.17504/protocols.io.bytapwie
https://www.protocols.io/view/crosslinking-assay-to-study-a-specific-cargo-coat-bytapwie
Javier Manzano-Lopez †, Sofia Rodriguez-Gallardo †, Susana Sabido-Bozo, Ana Maria Perez-Linero, Rafael Lucena, Antonio Cordones-Romero, Sergio Lopez, Auxiliadora Aguilera-Romero, Manuel Muñiz
TITLE: Crosslinking assay to study a specific cargo-coat interaction through a transmembrane receptor in the secretory pathway AUTHORS: Javier Manzano-Lopez †, Sofia Rodriguez-Gallardo †, Susana Sabido-Bozo, Ana Maria Perez-Linero, Rafael Lucena, Antonio C...
["[Yeast growth and culture] Transform the yeast strain genomically expressing Lst1-mCherry with a centromeric plasmid expressing GFP-tagged Gas1 under control of its own promoter (pRS416-GAS1-GFP).", "[Cell lysis] Quick thaw the cell pellets and immediately place on ice.", "[Cell lysis] Resuspend each cell pellet with...
63,519
Reverse transcription, primer pools preparation and multiplex PCR steps for DENV2 serotype
4
dx.doi.org/10.17504/protocols.io.4r3l2ob2pv1y/v1
https://www.protocols.click/view/reverse-transcription-primer-pools-preparation-and-b997r99n
Laís Ceschini, Carla Julia da Silva Pessoa Vieira, Gustavo Lima, Luisa Maria Inácio da Silva, Raul Emídio Lima, Tiago Graf, Gabriel Wallau
TITLE: Reverse transcription, primer pools preparation and multiplex PCR steps for DENV2 serotype AUTHORS: Laís Ceschini, Carla Julia da Silva Pessoa Vieira, Gustavo Lima, Luisa Maria Inácio da Silva, Raul Emídio Lima, Tiago Graf, Gabriel Wallau [DESCRIPTION] This step-by-step protocol describes the cDNA synthesis, pr...
["[Reverse transcription] Using a 2mL tube prepare the Mix 1 described below for 96 samples:\n\n \n \n \n Mix 1Reverse transcriptionVol. (1x)96 samples (+2 = 98 to keep some extra due to pipetting issues)Random Hexamers (50µM)1µL98µLdNTPs mix (10mM cada)1µL98µLTotal2µL194µL", "[Reverse transcription] Using 0,2mL PCR ...
58,078
Protocol of a systematic review with meta-analysis: The effects of physical exercise/activity on body composition of individuals with cardiometabolic multimorbidity
1
dx.doi.org/10.17504/protocols.io.b4x6qxre
https://www.protocols.io/view/protocol-of-a-systematic-review-with-meta-analysis-b4x6qxre
Juliene Gonçalves Costa, Igor Mariano, Victor Hugo Carrijo, Priccila Zuchinalli, Guilherme Morais Puga, Paula Aver Bretanha Ribeiro
TITLE: Protocol of a systematic review with meta-analysis: The effects of physical exercise/activity on body composition of individuals with cardiometabolic multimorbidity AUTHORS: Juliene Gonçalves Costa, Igor Mariano, Victor Hugo Carrijo, Priccila Zuchinalli, Guilherme Morais Puga, Paula Aver Bretanha Ribeiro ...
["[Background] Background \n\n Worldwide, more than 60% of the adult population has at least one chronic disease and 20% to 30% are affected by two or more chronic diseases, with cardiometabolic diseases being the most frequent and the main causes of death in the world. People with multiple chronic diseases have a...
37,956
HTAPP_TST- Nuclei isolation from frozen tissue
1
dx.doi.org/10.17504/protocols.io.bhbcj2iw
https://www.protocols.io/view/htapp-tst-nuclei-isolation-from-frozen-tissue-bhbcj2iw
Eugene Drokhlyansky, Nicholas Van Wittenberghe, Michal Slyper, Julia Waldman, Asa Segerstolpe, Orit Rozenblatt-Rosen, Aviv Regev
TITLE: HTAPP_TST- Nuclei isolation from frozen tissue AUTHORS: Eugene Drokhlyansky, Nicholas Van Wittenberghe, Michal Slyper, Julia Waldman, Asa Segerstolpe, Orit Rozenblatt-Rosen, Aviv Regev [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol describes a method based on work by </sp...
["[Nuclei isolation]\nUsing forceps, place tissue in the pre-filled well of the 6-well plate on ice (containing the 1 ml of TST; Step 1) and chop tissue for 10 minutes in the buffer using spring scissors.\n[On ice]", "[Nuclei isolation]\nWith a 1 ml pipette transfer the suspension from the 6-well plate to a 40 µm filte...
91,694
Deepwell Reusual Protocol
1
dx.doi.org/10.17504/protocols.io.j8nlko2x6v5r/v2
https://www.protocols.io/view/deepwell-reusual-protocol-c5sny6de
Morris Jack
TITLE: Deepwell Reusual Protocol AUTHORS: Morris Jack [DESCRIPTION] This is a protocol to reuse deepwells for sustainability practices [STEPS] SECTION: Growing cultures in a deep well 1. Cover a deep well with two layers of aluminium foil and tape the front and back to hold it securely. The tape also turns the alumi...
["[Growing cultures in a deep well] Cover a deep well with two layers of aluminium foil and tape the front and back to hold it securely. The tape also turns the aluminium foil into a lid for the deep well by temporarily peeling off the front tape.", "[Growing cultures in a deep well] Autoclave at 120C for 20min. After ...
103,679
In-Silico Validation of Biomarkers using ROC and AUC Curve Analysis in R: A Comprehensive Protocol
0
dx.doi.org/10.17504/protocols.io.q26g713qqgwz/v1
https://www.protocols.io/view/in-silico-validation-of-biomarkers-using-roc-and-a-dhg733zn
Adarsh V, Shreya Satyanarayan Bhat, Vidya Niranjan
TITLE: In-Silico Validation of Biomarkers using ROC and AUC Curve Analysis in R: A Comprehensive Protocol AUTHORS: Adarsh V, Shreya Satyanarayan Bhat, Vidya Niranjan [DESCRIPTION] Biomarkers are essential for the early detection, diagnosis, and management of diseases, particularly in complex conditions like Alzheimer'...
["[Introduction] Biomarkers play a crucial role in the early detection, diagnosis, and management of various diseases. However, the development and validation of reliable biomarkers is a complex and challenging process that requires rigorous evaluation of their analytical and clinical performance. Biomarker validation ...
43,696
PCR
4
dx.doi.org/10.17504/protocols.io.bnwqmfdw
https://www.protocols.io/view/pcr-bnwqmfdw
Zhujun Wei
TITLE: PCR AUTHORS: Zhujun Wei [STEPS] ?. Add the following reagent to a PCR tube.(50 μl). AB12×high Taq Master Mix (Enzyme)25 μl2Template2 μl3Forward Primer (10 μM)2 μl4Reverse Primer (10 μM)2 μl5ddH2O19 μl AB12×high Taq Master Mix (Enzyme)25 μl2Template2 μl3Forward Primer (10 μM)2 μl4Reverse Primer (10 μM)2 μl5ddH2...
["Add the following reagent to a PCR tube.(50 μl). AB12×high Taq Master Mix (Enzyme)25 μl2Template2 μl3Forward Primer (10 μM)2 μl4Reverse Primer (10 μM)2 μl5ddH2O19 μl\nAB12×high Taq Master Mix (Enzyme)25 μl2Template2 μl3Forward Primer (10 μM)2 μl4Reverse Primer (10 μM)2 μl5ddH2O19 μl", "Program the thermocycler as fo...
29,278
Minimal sample metadata for healthy liver tissue
null
dx.doi.org/10.17504/protocols.io.8t6hwre
null
Lenny Teytelman
TITLE: Minimal sample metadata for healthy liver tissue AUTHORS: Lenny Teytelman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is just a draft. Please feel free to revise and comment, but try to keep the total fields to a maximum of 20.</div></div> [STEPS]
[]
56,243
Total Starch Enzymatic Digestion
1
dx.doi.org/10.17504/protocols.io.b26tqhen
https://www.protocols.io/view/total-starch-enzymatic-digestion-b26tqhen
Lynn Doran, Amanda P. De Souza
TITLE: Total Starch Enzymatic Digestion AUTHORS: Lynn Doran, Amanda P. De Souza [DESCRIPTION] Enzymatic digestion of total soluble starch to glucose in plant tissue extracts for preparation for quantification via the GOD-POD Method (NZYtech). [BEFORE_START] Extract and dry total starch pellet from plant tissue p...
["Prepare fresh daily 120 U/mL α-amylase in MOPS buffer. 1 mL per sample will be needed. Initial concentration of α-amylase is 1000 U/mL. Use C1V1 = C2V2 to calculate the volume of α-amylase and MOPS buffer to use.", "Prepare fresh daily 30 U/mL amyloglucosidase in acetate buffer. 1 mL per sample will be needed. Initi...
77,893
Characterization of the VKORC1 and CYP2C9 genotypes
1
dx.doi.org/10.17504/protocols.io.kxygx9edzg8j/v3
https://www.protocols.io/view/characterization-of-the-vkorc1-and-cyp2c9-genotype-cqbdvsi6
Mirsada Causevic, Edin Begic
TITLE: Characterization of the VKORC1 and CYP2C9 genotypes AUTHORS: Mirsada Causevic, Edin Begic [DESCRIPTION] Vitamin K antagonists (e.g. warfarin) are anticoagulants which represent widely prescribed drugs for prevention and treatment of thromboembolic disorders. Warfarin's molecular target is vitamin K epoxide re...
["[Genomic DNA extraction] Patients' whole blood was collected in ethylenediaminetetraacetic acid (EDTA)-containing tubes and stored at -20°C until use. Genomic DNA extraction from the human whole blood, that is, leukocytes, was carried out according to the protocol described by Subbarayan PR and colleagues (doi: 10.21...
53,570
Conducting Online Research With Infants
1
null
https://www.protocols.io/view/conducting-online-research-with-infants-byjapuie
ajthomas
TITLE: Conducting Online Research With Infants AUTHORS: ajthomas [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a step-by-step process for running synchronous online studies with infants: studies in which you meet parents and infants via video chat. This method is meant to be very similar ...
["[Stimuli Set Up]\nMake videos of stimuli by using keynote or powerpoint. Once you have the stimuli you want to use. You can use keynote or powerpoint to make a long video that includes attention getters etc. Make your keynote file that includes videos and attention getters in the order you want them. Then select File...
98,082
Spatial N-glycomics with MALDI-MSI for human kidney tissue
1
dx.doi.org/10.17504/protocols.io.8epv5j1m4l1b/v3
https://www.protocols.io/view/spatial-n-glycomics-with-maldi-msi-for-human-kidn-db2a2qae
Dusan Velickovic, Kumar Sharma, Theodore Alexandrov, Chris Anderton
TITLE: Spatial N-glycomics with MALDI-MSI for human kidney tissue AUTHORS: Dusan Velickovic, Kumar Sharma, Theodore Alexandrov, Chris Anderton [DESCRIPTION] This protocol describes the procedure to obtain high quality MALDI mass spectrometry images of N-linked glycans from formalin-fixed paraffin embedded tissue. Thi...
["[Scope] This protocol describes the procedure to obtain high quality MALDI mass spectrometry images of N-linked glycans from formalin-fixed paraffin embedded tissue.", "[Health and Safety] Wear nitrile gloves and safety glasses. Follow standard laboratory safety procedures.", "[Equipment] Equipment Required:", "[Equi...
88,667
Village Nuclei Isolation With Myelin Removal
4
dx.doi.org/10.17504/protocols.io.n2bvj3dxblk5/v2
https://www.protocols.io/view/village-nuclei-isolation-with-myelin-removal-c2t3yeqn
Steve McCarroll, Emi Ling, Melissa Goldman, Nora Reed
TITLE: Village Nuclei Isolation With Myelin Removal AUTHORS: Steve McCarroll, Emi Ling, Melissa Goldman, Nora Reed [DESCRIPTION] Isolation of nuclei from fresh-frozen brain tissue from sets of multiple (typically 2-20) human donors for analysis as a “cell village” (Wells et al., PMID 36796362) in which nuclei from all...
["[Before Starting] Gather Supplies\nRazor Blades\nGlass slides\nSyringes with needles (3 mL syringe with 26 1⁄2 gauge needle)\nMyelin removal beads (cat # 130-096-731) https://www.miltenyibiotec.com/US-en/products/myelin-removal-beads-ii-human-mouse-rat.html#gref \nEppendorf tubes (1.5 mL and 5 mL)\nEppendorf or Raini...
28,384
MojoSort™ Isolation Kits Protocol - 4
null
dx.doi.org/10.17504/protocols.io.7x8hprw
null
Sam Li
TITLE: MojoSort™ Isolation Kits Protocol - 4 AUTHORS: Sam Li [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Product description and procedure summary:</span></div><div class = "text-block"><span>Target cells are depleted by incubating your sample with the biotin an...
["Prepare cells from your tissue of interest or blood without lysing erythrocytes. Kits for human samples have been optimized for PBMCs, please prepare the cells using a suitable method.", "In the final wash of your sample preparation, resuspend the cells in MojoSort™ Buffer by adding up to 4mL in a 5 mL (12 x 75 mm) p...
21,670
Chlamydia trachomatis PCR
null
dx.doi.org/10.17504/protocols.io.zeef3be
null
Ana Ximena Kiguen, Jessica Paola Mosmann, Raul Fernando Venezuela, Cecilia Gabriela Cuffini
TITLE: Chlamydia trachomatis PCR AUTHORS: Ana Ximena Kiguen, Jessica Paola Mosmann, Raul Fernando Venezuela, Cecilia Gabriela Cuffini [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:justify"><span>OmpA gene PCR: PCR DNA extract (5 μl) was used to amplify a ...
["Pre Amplification Mix( NRO/NLO for ompA gene, and CTP1/CTP2 for cryptic Plasmid)\n[Mint Master Mix]\n[Agua ultrapura]\n[each primer]", "Add Extracted DNA\n[DNA]", "In a thermocycler , incubate as follows for ompA gene:49 amplification cycles as follows:Incubate as follows for Cryptic Plasmid:35 amplification cycles a...
28,962
Viral Preparation
null
dx.doi.org/10.17504/protocols.io.8iahuae
null
Aditya Mohan
TITLE: Viral Preparation AUTHORS: Aditya Mohan [STEPS] ?. [Day 1] Coat the plates with 1% Poly-L-Lysine solution and leave in dry incubator overnight. ?. [Day 2 ] Plate 6E6 HEK293 T cells overnight in DMEM media ?. [Day 3] In 2 1.5 mL epindorph tube add 150 uL of OptiMEM. In one of them add 15 uL of lipofectamine. In ...
["[Day 1]\nCoat the plates with 1% Poly-L-Lysine solution and leave in dry incubator overnight.", "[Day 2 ]\nPlate 6E6 HEK293 T cells overnight in DMEM media", "[Day 3]\nIn 2 1.5 mL epindorph tube add 150 uL of OptiMEM. In one of them add 15 uL of lipofectamine. In the other add 10 uL of packaging plasmid and 10 uL of ...
81,048
Size selection (Purification)
1
dx.doi.org/10.17504/protocols.io.bp2l695eklqe/v1
https://www.protocols.io/view/size-selection-purification-ctdywi7w
Tsu-Chun Hung
TITLE: Size selection (Purification) AUTHORS: Tsu-Chun Hung [DESCRIPTION] Size selection (Purification) [STEPS] 1. Prepare 20 µL sample and 9 µL magnetic beads in 1.5 mL eppendorf tube. 2. Mix sample and beads gently by flicking then flash spin the tube. Put on the regular rack for 5mins 5 min. 3. Transfer the...
["Prepare 20 µL sample and 9 µL magnetic beads in 1.5 mL eppendorf tube.", "Mix sample and beads gently by flicking then flash spin the tube. Put on the regular rack for 5mins 5 min.", "Transfer the tube to the magnetic rack. After most of the magnetic beads attach to the wall, remove the supernant.", "Add 300 µL 75% e...
45,161
3.1 Preparation of Monocellular Lung Suspension from a Mouse Lung Lobe
4
dx.doi.org/10.17504/protocols.io.bqchmst6
https://www.protocols.io/view/3-1-preparation-of-monocellular-lung-suspension-fr-bqchmst6
Helen Graves, Steven Evans, Michael Fauler, Manfred Frick, Sterghios A. Moschos
TITLE: 3.1 Preparation of Monocellular Lung Suspension from a Mouse Lung Lobe AUTHORS: Helen Graves, Steven Evans, Michael Fauler, Manfred Frick, Sterghios A. Moschos [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>The clinical potential of DNA and RNA-targeting therapeutics for airways diseas...
["[3.1 Preparation of Monocellular Lung Suspension from a Mouse Lung Lobe]\nRinse the fresh, perfused mouse lung, with sterile PBS to eliminate any blood.", "[3.1 Preparation of Monocellular Lung Suspension from a Mouse Lung Lobe]\nTransfer the lung lobe to in a sterile tissue vial (ensure full tissue submersion) and ...
43,142
Stranded Transcript Count Table Generation from Long Reads
1
dx.doi.org/10.17504/protocols.io.bndema3e
https://www.protocols.io/view/stranded-transcript-count-table-generation-from-lo-bndema3e
David Eccles
TITLE: Stranded Transcript Count Table Generation from Long Reads AUTHORS: David Eccles [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for comparing different samples at the transcript level, using long reads that are mapped to transcripts.</div><div class = "text-block"><span styl...
["[Demultiplex Reads ]\nDemultiplex and orient reads as per the protocol Preparing Reads for Stranded Mapping. It is expected that these demultiplexed reads will be split up in the current directory, and coupled with a 'barcode_counts.txt' file. If that's the case, the following should work:for bc in $(awk '{print $...
61,694
Antibiotic Sensitivity Assay for Heterometopus palaeformis (strain RAJCA)
4
dx.doi.org/10.17504/protocols.io.x54v9y3mzg3e/v1
https://www.protocols.io/view/antibiotic-sensitivity-assay-for-heterometopus-pal-b8g6rtze
Fatma Gomaa, Johana Rotterova, Roxanne Berinate, Maria Pachiadaki, Virginia Edgcomb
TITLE: Antibiotic Sensitivity Assay for Heterometopus palaeformis (strain RAJCA) AUTHORS: Fatma Gomaa, Johana Rotterova, Roxanne Berinate, Maria Pachiadaki, Virginia Edgcomb [DESCRIPTION] Goal: To determine the optimal concentration of three antibiotics, Puromycin, Geneticin (G418) and Blasticidin, that will effec...
[]
103,236
SANGER Sequencing EFGL
0
dx.doi.org/10.17504/protocols.io.yxmvmew25g3p/v1
https://www.protocols.io/view/sanger-sequencing-efgl-dg3c3yiw
EagleFish GeneticsLab
TITLE: SANGER Sequencing EFGL AUTHORS: EagleFish GeneticsLab [DESCRIPTION] This protocol describes how the Eagle Fish Genetics Lab (EFGL) prepares extracted DNA samples for the ABI 3500xL Genetic Analyzer, and how we run and collect data from this machine. This process is achieved by PCR amplification, ExoSAP-IT purif...
["[PCR Amplification of Target Sequence] Materials needed: \n• Forward and reverse primers\n• Heat Seal\n• 1.5mL vials \n• PCR master mix reagents/kits \n• DNA tray(s) \n• Unskirted PCR plate(s)\n• Pipette and tip", "[SECTION 1 – PCR Purification with ExoSAP-IT] Materials needed: \n• ExoSAP-IT Express reagent (pre-al...
86,328
Reverse transcription, primer pools preparation and multiplex PCR steps for DENV1 serotype for genomic sequencing
4
dx.doi.org/10.17504/protocols.io.5jyl8pjyrg2w/v1
https://www.protocols.io/view/reverse-transcription-primer-pools-preparation-and-cyiyxufw
Laís Ceschini, Carla Julia da Silva Pessoa Vieira, Luisa Maria Inácio da Silva, Gustavo Lima, Raul Emídio, Tiago Graf, Gabriel Luz Wallau
TITLE: Reverse transcription, primer pools preparation and multiplex PCR steps for DENV1 serotype for genomic sequencing AUTHORS: Laís Ceschini, Carla Julia da Silva Pessoa Vieira, Luisa Maria Inácio da Silva, Gustavo Lima, Raul Emídio, Tiago Graf, Gabriel Luz Wallau [DESCRIPTION] This step-by-step protocol describes ...
["[Reverse transcription] Using a 2mL tube prepare the Mix 1 described below for 96 samples:", "[Reverse transcription] Using 0,2mL PCR tubes or 96 wells plates add 11-16µL of extracted RNA from\nRT-PCR positive samples. Add 2µL of Mix 1 to the tube/well and take it to the\nthermocycler with the following set up\n\n65º...
28,807
Lentiviral transduction of iPSCs with sgRNAs and sgRNA libraries
null
dx.doi.org/10.17504/protocols.io.8dfhs3n
null
Ruilin Tian, Jason Hong, Sydney Sattler, Martin Kampmann
TITLE: Lentiviral transduction of iPSCs with sgRNAs and sgRNA libraries AUTHORS: Ruilin Tian, Jason Hong, Sydney Sattler, Martin Kampmann [STEPS] ?. [Day 0: Seeding] 18 – 24 hours before transfection, seed 293T cells into a 6 well plate or other format with a density that will make the cells 80 – 95 % confluent on the...
["[Day 0: Seeding]\n18 – 24 hours before transfection, seed 293T cells into a 6 well plate or other format with a density that will make the cells 80 – 95 % confluent on the day of transfection. Refer to a seeding chart if necessary to seed appropriate density.", "[Day 0: Seeding]\nIncubate overnight.", "[Day 1: Transf...
90,731
Isothermal Titration Calorimetry of the Rubicon RH domain and Rab7
4
dx.doi.org/10.17504/protocols.io.q26g7pwd3gwz/v1
https://www.protocols.io/view/isothermal-titration-calorimetry-of-the-rubicon-rh-c4ujywun
Dan Tudorica
TITLE: Isothermal Titration Calorimetry of the Rubicon RH domain and Rab7 AUTHORS: Dan Tudorica [DESCRIPTION] 1:1 binding of Rubicon and Rab7 [STEPS] SECTION: Protein prep 1. On day prior to experiment, codialyze both binding partners to use in the the same beaker. SECTION: Protein prep 2. Prepare 2 L of 50 mM HEPES ...
["[Protein prep] On day prior to experiment, codialyze both binding partners to use in the the same beaker.", "[Protein prep] Prepare 2 L of 50 mM HEPES 7.5, 150 mM NaCl, 10 mM TCEP, and degas thoroughly at room temperature under vacuum with stir bar agitation, then chill in fridge until ready for dialysis.", "[Protein...
null
null
null
dx.doi.org/10.17504/protocols.io.ftybnpw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>A protocol used to generate a PDMS microfluidic chip. Work was funded by Cambridge Synthetic Biology Strategic Research Initative (SRI) SynBio Fund.</p> <p>http://www.synbio.cam.ac.uk/synbiofund</p> <p> </p> [BEFORE_START] <ol> <li><span style="line-height: 16px;">Use AutoDe...
[]
24,123
Biochemical Measures of Neuropathy - Western Blot Stripping
null
dx.doi.org/10.17504/protocols.io.3s3gngn
null
Eva Feldman
TITLE: Biochemical Measures of Neuropathy - Western Blot Stripping AUTHORS: Eva Feldman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block">Oxidative stress is highly correlated with the metabolic changes caused by hypergly...
["[Performing assay:]\nRainbow markers do not withstand stripping, so if you don’t have a biotinylated marker on your blot, be sure to mark the location of the rainbow markers with a pen or pencil before stripping 1. Incubate your blot for 10-20 min at 70ºC. to strip. (10 min. for 20-40 µg protein and 15-20 min. for lo...
83,400
S. aureus biofilm removal multi-assay
6
dx.doi.org/10.17504/protocols.io.bp2l6xk6klqe/v1
https://www.protocols.click/view/s-aureus-biofilm-removal-multi-assay-cvpgw5jw
Tomasz Swebocki
TITLE: S. aureus biofilm removal multi-assay AUTHORS: Tomasz Swebocki [DESCRIPTION] The following protocol describes culture and treatment of the S.aureus-based biofilm and techniques that can be used to asses its eradication/removal. The protocol covers basic preparation and three techniques, namely: growth control, ...
["[Biofilm culture] Culture bacteria strains in TSB at 35 °C for approximately 240 min in order to reach mid-log phase (OD600 0.5-0.6).", "[Biofilm culture] Pellet the cells by centrifugation (4500 rpm, 5 min, 20 °C) then wash twice with potassium phosphate buffer (PPB; 100 millimolar (mM), pH 7).\n\nFor washing, redis...
null
null
null
dx.doi.org/10.17504/protocols.io.ihncb5e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Please contact Dr. Steven Wilhelm (wilhelm@utk.edu) for additional information regarding this protocol.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
50,982
Cross-linking/MS-analysis of Thr72-phosphorylated Rab8A and PPM1H (D288A) complex
1
dx.doi.org/10.17504/protocols.io.bv2en8be
https://www.protocols.io/view/cross-linking-ms-analysis-of-thr72-phosphorylated-bv2en8be
Pawel Lis, Pui Yiu Lam, Axel Knebel , Kerryn Berndsen, Dario Alessi
TITLE: Cross-linking/MS-analysis of Thr72-phosphorylated Rab8A and PPM1H (D288A) complex AUTHORS: Pawel Lis, Pui Yiu Lam, Axel Knebel , Kerryn Berndsen, Dario Alessi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A subset of Rab proteins, including Rab8A, have been identified as substrates of the L...
["[Protein buffer exchange]\nPrepare a beaker containing of HEPES , NaCl, MgCl2 buffer and a magnetic stir bar. Set the beaker on a magnetic stirrer with gentle stirring, at .\nAs the cross-linking reaction can be inhibited by amine containing buffers, a buffer exchange step for both, PPM1H and pRab8A protein solut...
73,800
Eggs injections
4
null
https://www.protocols.io/view/eggs-injections-ckbgusjw
FishFloorUCL
TITLE: Eggs injections AUTHORS: FishFloorUCL [DESCRIPTION] Protocol for microinjections of 0-dpf eggs @FishFloorUCL. [STEPS] SECTION: Set-up breeding pairs 1. Set up the breeding pairs the night before or order them from the fish facility. Arrive early in the morning, 9AM for single cell, 9:30 for fish facility injec...
["[Set-up breeding pairs] Set up the breeding pairs the night before or order them from the fish facility. Arrive early in the morning, 9AM for single cell, 9:30 for fish facility injections.\n\nMake sure to book the injector in advance.", "[Calibrate the needle] Open the red valve for your injector, you should see the...
102,735
Chromogenic in situ hybridisation
4
null
https://www.protocols.io/view/chromogenic-in-situ-hybridisation-dgjp3umn
Stephen Carter
TITLE: Chromogenic in situ hybridisation AUTHORS: Stephen Carter [DESCRIPTION] The protocol for performing chromogenic in situ hybridisations in zebrafish embryos and larvae in the Wilson lab. [STEPS] SECTION: Probe synthesis 2. If the template DNA is a plasmid, it must first be linearised by restriction enzyme dige...
["[Probe synthesis] If the template DNA is a plasmid, it must first be linearised by restriction enzyme digestion. If it is a PCR product, it can be used directly. \n\nPrepare probe synthesis reaction mix\n\nTemplate DNA (PCR product or plasmid) -0.5-1 µg\n10x DIG RNA mix (Roche) - 2 µL\n10x transcription buffer - 2 µL...
98,415
The Parkinson’s Progression Markers Initiative (PPMI) Clinical - Establishing a Deeply Phenotyped PD Cohort AM 3.2
1
dx.doi.org/10.17504/protocols.io.n92ldmw6ol5b/v2
https://www.protocols.io/view/the-parkinson-s-progression-markers-initiative-ppm-dccp2svn
Kenneth Marek
TITLE: The Parkinson’s Progression Markers Initiative (PPMI) Clinical - Establishing a Deeply Phenotyped PD Cohort AM 3.2 AUTHORS: Kenneth Marek [DESCRIPTION] This protocol details the Parkinson’s Progression Markers Initiative (PPMI) Clinical - establishing a deeply phenotyped PD cohort. [GUIDELINES] APPENDIX 1 – He...
["[PURPOSE OF STUDY] Primary Objectives of PPMI Clinical:\nThe primary objectives include to:", "[PURPOSE OF STUDY] Establish standardized protocols for acquisition, transfer and analysis of clinical, digital, imaging, biologic and genetic data that can be used by the PD research community. This protocol will build on ...
37,343
Targeted ExSeq -- Sequencing Library Preparation
1
dx.doi.org/10.17504/protocols.io.bgp7jvrn
https://www.protocols.io/view/targeted-exseq-sequencing-library-preparation-bgp7jvrn
Anubhav Sinha, Asmamaw T. Wassie, Fei Chen, Yi Cui, Ed Boyden
TITLE: Targeted ExSeq -- Sequencing Library Preparation AUTHORS: Anubhav Sinha, Asmamaw T. Wassie, Fei Chen, Yi Cui, Ed Boyden [DESCRIPTION] This protocol accompanies Expansion Sequencing (ExSeq), describing the process of targeted ExSeq library preparation for a sample that has been processed according ...
["[Padlock Probe Hybridization] Padlock Probe Hybridization\n\nThe first step of library preparation is to hybridize padlock probes to the RNA transcripts of interest within the expanded samples.", "[Universal Amplicon Detection Hybridization] Amplicon Detection Hybridization\n\nA rapid quality control assay for the li...
98,529
Carrier-assisted One-pot Sample Preparation for Targeted Proteomics Analysis of Small Numbers of Human Cells
4
dx.doi.org/10.17504/protocols.io.4r3l24zbjg1y/v2
https://www.protocols.io/view/carrier-assisted-one-pot-sample-preparation-for-ta-dcf92tr6
Kendall Martin, Tong Zhang, William B. Chrisler, Fillmore L. Thomas, WEI-JUN QIAN, Tujin Shi
TITLE: Carrier-assisted One-pot Sample Preparation for Targeted Proteomics Analysis of Small Numbers of Human Cells AUTHORS: Kendall Martin, Tong Zhang, William B. Chrisler, Fillmore L. Thomas, WEI-JUN QIAN, Tujin Shi [DESCRIPTION] Protein analysis of small numbers of human cells is primarily achieved by targeted prot...
["[PROCEDURE] Pretreatment of PCR tubes", "[PROCEDURE] Add 100 µL of nonhuman (e.g., Shewanella oneidensis)cell lysate digests at 0.2 µg/µL to PCR tubes. Incubate at room temperature for overnight to coat PCR tube surface.", "[PROCEDURE] Remove the cell lysate digests by pipetting, rinse PCR tubes with HPLC-grade water...
null
null
null
dx.doi.org/10.17504/protocols.io.pmxdk7n
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
107,243
A Protocol for Assessing Open Data Practices: Honours Students Can Lead the Way.
0
dx.doi.org/10.17504/protocols.io.kxygxyxmdl8j/v2
https://www.protocols.io/view/a-protocol-for-assessing-open-data-practices-honou-dkyj4xun
Haya Deeb, Tomasz Zieliński, Andrew J. Millar
TITLE: A Protocol for Assessing Open Data Practices: Honours Students Can Lead the Way. AUTHORS: Haya Deeb, Tomasz Zieliński, Andrew J. Millar [DESCRIPTION] Introduction  The culture of scientific research is increasingly recognising the importance of Open Data. Open Data practices involve making research data freely ...
["[Sampling Framework and Selection Process] Research groups within the biosciences at the University of Edinburgh were selected for study based on the educational interests of the undergraduate honours student researchers. Articles from each group were retrieved from the University’s public Edinburgh Research Explorer...
95,715
MycoEd Lab: CU Denver Protocols
0
dx.doi.org/10.17504/protocols.io.n2bvj3k6xlk5/v1
https://www.protocols.io/view/mycoed-lab-cu-denver-protocols-c9qbz5sn
Andrew W. Wilson, sara.branco
TITLE: MycoEd Lab: CU Denver Protocols AUTHORS: Andrew W. Wilson, sara.branco [DESCRIPTION] These lab protocols were written for the Fall 2023 Mycology course at University of Colorado Denver, Department of Integrative Biology instructed by Sara Branco and Andrew Wilson. The course is designed to guide students throug...
[]
65,237
Oprah Winfrey Keto Gummies 
1
dx.doi.org/10.17504/protocols.io.dm6gpb13jlzp/v1
https://www.protocols.io/view/oprah-winfrey-keto-gummies-cbxvspn6
jamshruckz
TITLE: Oprah Winfrey Keto Gummies  AUTHORS: jamshruckz [DESCRIPTION] Progressing to ketosis is certainly not a speedy cycle. It requires investment for your body to become accustomed to this better approach for living. Certain individuals experience a ton of desires, weariness, and other upsetting incidental effects...
["➢Official Link:- https://www.facebook.com/OprahWinfreyGummies/\n➢ Product Name —Oprah Winfrey Keto Gummies\n➢ Composition—NATURAL\n➢ Side-Effects— NA\n➢ Availability— Online\n➢ Rating— ⭐⭐⭐⭐⭐\n➢ Official Website (Sale Is Live) —\nShop Now:- https://topcbdoilmart.com/oprah-winfrey-keto-gummies/\n\nOprah Winfrey Keto Gu...
39,056
teste
1
dx.doi.org/10.17504/protocols.io.bidqka5w
https://www.protocols.io/view/teste-bidqka5w
Nancy Sotero
TITLE: teste AUTHORS: Nancy Sotero [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">teste teste teste</div></div> [STEPS] ?. testei ?. testei
["testei", "testei"]
27,848
Tranformation of Thalassiosira pseudonana via bacterial conjugation
null
dx.doi.org/10.17504/protocols.io.7fghjjw
null
Ana Cristina Jaramillo Madrid, Justin Ashworth
TITLE: Tranformation of Thalassiosira pseudonana via bacterial conjugation AUTHORS: Ana Cristina Jaramillo Madrid, Justin Ashworth [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>This protocol has been successfully used to express nourseothricin resistance gene, mVenus fluorescence protein and...
["[Growth and preparation of E. coli donor]\nInoculate 5 mL LB medium (gentamicin+antibiotic 2) with bacterial colonies from the gentamicin +antibiotic 2 plates. Grow overnight.", "[Growth and preparation of E. coli donor]\nStart a 150 mL LB subculture with the 5 mL overnight culture (recommended starting OD600 either ...
61,760
Final QC, Pooling and Sequencing 
4
dx.doi.org/10.17504/protocols.io.yxmvmnw29g3p/v1
https://www.protocols.io/view/final-qc-pooling-and-sequencing-b8i8ruhw
Katarina A Cohen, Khai-Minh H Nguyen, Oksana Polesskaya, Abraham Palmer
TITLE: Final QC, Pooling and Sequencing  AUTHORS: Katarina A Cohen, Khai-Minh H Nguyen, Oksana Polesskaya, Abraham Palmer [DESCRIPTION] This protocol is conducted after a set of libraries are completed and ready to quantify and pool. This protocol outlines the final steps before submitting for sequencing. [BEFORE_ST...
["[Library QC] Quantify purity and concentration of library with Nanodrop and a Qubit Assay\nObtain average fragment size of library with Tapestation (D1000 Assay)", "[Pooling] Download", "[Pooling] Increase \"Target Vol (uL) per Sample\" if any Sample Vol is lower than 1ul\nUse when adding water to final pool", "[Ch...
86,984
Tissue clearing of human cardiac tissues using modified iDISCO+ protocol
4
dx.doi.org/10.17504/protocols.io.yxmvm3eo9l3p/v1
https://www.protocols.io/view/tissue-clearing-of-human-cardiac-tissues-using-mod-cy7gxzjw
Peter Hanna, Kalyanam Shivkumar, Shumpei Mori, Olujimi Ajijola
TITLE: Tissue clearing of human cardiac tissues using modified iDISCO+ protocol AUTHORS: Peter Hanna, Kalyanam Shivkumar, Shumpei Mori, Olujimi Ajijola [DESCRIPTION] The purpose of this protocol is to evaluate the myocardial innervation of the human heart. Using donor hearts rejected for human transplantation, imaging...
["[Tissue Fixation] Donor human hearts are obtained from the organ procurement organization and undergo perfusion fixation.", "[Tissue Fixation] Suture three of the pulmonary veins and the inferior vena cava closed. One pulmonary vein and the superior vena cava are left patent as outflow of perfusate.", "[Tissue Fixati...
72,742
High-throughput Assay for Screening Fungal Isolates against Polyphenolic Compounds 
4
null
https://www.protocols.io/view/high-throughput-assay-for-screening-fungal-isolate-cjaeuibe
Jana M U'Ren, Megan Nickerson
TITLE: High-throughput Assay for Screening Fungal Isolates against Polyphenolic Compounds  AUTHORS: Jana M U'Ren, Megan Nickerson [DESCRIPTION] This protocol was developed for the screening of fungal isolates against polyphenolic compounds to test their capacity for detoxification. [STEPS] SECTION: Media and Solutio...
["[Media and Solution Preparation- Aspergillus nidulans Defined Media plus Carrageenan] Combine:\n- 900mL nanopure water\n-50mL 20X Sodium nitrate salts\n-1mL Trace elements", "[Media and Solution Preparation- Aspergillus nidulans Defined Media plus Carrageenan] Bring the pH to 6.5.", "[Media and Solution Preparation- ...
null
null
null
dx.doi.org/10.17504/protocols.io.c4gytv
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Must be made fresh before experiment because of the Sucrose. <br />For 20 mL solutions. [BEFORE_START] Prepare solution with sterile milliQ water and filter sterilize or autoclave before use. [GUIDELINES] Final Concentration of Lysis Buffer<br />40 mM EDTA<br />50 mM Tris (pH ...
[]
97,849
Solid Growth Medium - Yeast
0
dx.doi.org/10.17504/protocols.io.j8nlk85x5l5r/v1
https://www.protocols.io/view/solid-growth-medium-yeast-dbsz2nf6
Mathias Hammer, Ammeret Rossouw, Azra Lari, Ben Montpetit, David Grunwald
TITLE: Solid Growth Medium - Yeast AUTHORS: Mathias Hammer, Ammeret Rossouw, Azra Lari, Ben Montpetit, David Grunwald [DESCRIPTION] This protocol describes the steps to prepare solid culture medium for Saccharomyces cerevisiae. [BEFORE_START] Have the following solutions premixed: Glucose 20% 500 ml solution: Concen...
["[Preparation of ~20 agar plates of yeast growth medium] Compound medium for autoclave", "[Preparation of ~20 agar plates of yeast growth medium] Fill a 500 ml flask with 449 mLddH2O. \nAdd a magnetic stirring bar and place the flask on a stirring hot plate.", "[Preparation of ~20 agar plates of yeast growth medium] A...
49,629
Pneumonia Associated with Invasive and Noninvasive Oxygenation Strategies for Acute Hypoxemic Respiratory Failure in Adults: A Systematic Review and Network Meta-analysis Protocol
1
dx.doi.org/10.17504/protocols.io.bup5nvq6
https://www.protocols.io/view/pneumonia-associated-with-invasive-and-noninvasive-bup5nvq6
Satoshi Hokari, Shunsuke Kimata, Masaaki Sakuraya, Hiromu Okano, Tomoyuki Masuyama
TITLE: Pneumonia Associated with Invasive and Noninvasive Oxygenation Strategies for Acute Hypoxemic Respiratory Failure in Adults: A Systematic Review and Network Meta-analysis Protocol AUTHORS: Satoshi Hokari, Shunsuke Kimata, Masaaki Sakuraya, Hiromu Okano, Tomoyuki Masuyama [DESCRIPTION] <div class = "text-blocks"...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.h2db8a6
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
71,559
Interventions promoting physical activity among adolescents in Sub-Saharan Africa: a scoping review protocol
1
dx.doi.org/10.17504/protocols.io.n92ldpzpnl5b/v1
https://www.protocols.io/view/interventions-promoting-physical-activity-among-ad-ch5ft83n
Cécile Longchamps, Oumar Bassoum, Adama Faye, Valéry Ridde
TITLE: Interventions promoting physical activity among adolescents in Sub-Saharan Africa: a scoping review protocol AUTHORS: Cécile Longchamps, Oumar Bassoum, Adama Faye, Valéry Ridde [DESCRIPTION] Objective: To describe the existing health promotion interventions that aim to promote physical activity among adolescent...
["[Introduction] Non-communicable diseases (NCDs) cause 41 million deaths annually in the world, representing 71% of all deaths, and more than three quarters of these deaths take place in lower-middle-income countries (LMIC)1. The African region is undergoing an epidemiological transition with a double burden of commun...
null
null
null
dx.doi.org/10.17504/protocols.io.e7gbhjw
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Picking colonies from TCBS Agar plates to 96 well plates for growth</p> [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
21,294
Quantifying Fluorescent Cells in Mammalian Cell Tissue Culture
null
dx.doi.org/10.17504/protocols.io.y2nfyde
null
Harley King
TITLE: Quantifying Fluorescent Cells in Mammalian Cell Tissue Culture AUTHORS: Harley King [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Quantifying the change in fluorescent status of cells over time can be an important data point in experimental analysis. Optimally, quantification can be accomp...
["[Preparation and Transfection of Cells]\nA six-well plate is seeded with cells at a typical pre-transfection density. The following day cells are transfected.", "[Capture Plate Images]\nImages can be taken of the plate 24 twenty-four hours post transfection (hpt). Depending on the vector and the promoter, fluorescenc...
107,688
DDNS Data analysis and quality control checks
0
dx.doi.org/10.17504/protocols.io.5qpvok4jxl4o/v1
https://www.protocols.io/view/ddns-data-analysis-and-quality-control-checks-dmeg43bw
Alex Shaw, Joyce Akello, Catherine Troman, Aine OToole, c.ansley, Catherine Pratt, Erika Bujaki, Zoe Vance, rachel.colquhoun, Andrew Rambaut, Javier Martin, Nick Grassly
TITLE: DDNS Data analysis and quality control checks AUTHORS: Alex Shaw, Joyce Akello, Catherine Troman, Aine OToole, c.ansley, Catherine Pratt, Erika Bujaki, Zoe Vance, rachel.colquhoun, Andrew Rambaut, Javier Martin, Nick Grassly [DESCRIPTION] This standard operating procedure indicates how to perform data analysis...
["[Post Sequencing Run Checks] Perform the post sequencing run checks by confirming the following points manually\n a. Did the sequencing run complete its full run duration (check the\nMinKNOW run report). \n b. Was there no sudden reduction in pore numbers i.e. pores numbers did not fall beneath 400 (or 25% of total p...
64,975
In situ high-speed brightfield imaging for studies of aquatic organisms
4
dx.doi.org/10.17504/protocols.io.kxygxz4ykv8j/v2
https://www.protocols.io/view/in-situ-high-speed-brightfield-imaging-for-studies-cbppsmmn
Sean P. Colin, Brad J. Gemmell, John H. Costello, Kelly R R Sutherland
TITLE: In situ high-speed brightfield imaging for studies of aquatic organisms AUTHORS: Sean P. Colin, Brad J. Gemmell, John H. Costello, Kelly R R Sutherland [DESCRIPTION] Behavioral measurements of fragile aquatic organisms require specialized in situ techniques.We developed an in situ brightfield camera set-up ...
["[Select field site] This system is lightweight, compact and portable and can be used SCUBA diving from shore, docks or boats.The camera system can also potentially be towed vertically from a research vessel.", "[Assemble equipment] The brightfield camera system relies entirely on available off the shelf components an...
24,068
Thawing and Seeding Frozen Cells
null
dx.doi.org/10.17504/protocols.io.3rcgm2w
null
Kenneth Schackart
TITLE: Thawing and Seeding Frozen Cells AUTHORS: Kenneth Schackart [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">How to thaw cells from the liquid nitrogen storage and seed into a tissue culture flask</div></div> [STEPS] ?. [Thaw Cells] Thaw cells by suspending cryotube in water bath until compl...
["[Thaw Cells]\nThaw cells by suspending cryotube in water bath until completely thawed, but no longer than necessary", "[Transfer cell suspension]\nWithin biosafety cabinet, transfer cell suspension to 15 mL centrifuge tube using 1000 μL pipette.", "[Dilute freezing medium]\nAdd warmed cell culture medium to cell su...
41,590
Cloning of Bacillus mycoides
1
dx.doi.org/10.17504/protocols.io.bkuwkwxe
https://www.protocols.io/view/cloning-of-bacillus-mycoides-bkuwkwxe
Andreea S
TITLE: Cloning of Bacillus mycoides AUTHORS: Andreea S [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left"><span style = "font-weight:bold;">Introducing NLP14a in the genome of B. mycoides: </span><span> In order to create plasmid pYCR-gamyNLP, the backb...
["[Making electro competent cells]\nPick 1 colony of B. mycoides M2E_15 and inoculate it in BHIS", "[Making electro competent cells]\nWhen OD600nm reached 0.85 add 2% glycine and 2% threonine in order to weaken the cell wall.", "[Making electro competent cells]\nGrow the cells overnight at\n30 33", "[Making electro com...
82,448
Regular maintenance of human pluripotent stem cells
4
dx.doi.org/10.17504/protocols.io.bp2l69obdlqe/v1
https://www.protocols.click/view/regular-maintenance-of-human-pluripotent-stem-cell-curqwv5w
Narayana Yadavalli, Shawn M. Ferguson
TITLE: Regular maintenance of human pluripotent stem cells AUTHORS: Narayana Yadavalli, Shawn M. Ferguson [DESCRIPTION] This protocol describes the regular maintenance and passaging human iPSCs. [STEPS] SECTION: Day 1 1. Pre coat a 6 well dish with Matrigel matrix for 1440 min or 1 hour. SECTION: Day 1 2. Thaw the f...
["[Day 1] Pre coat a 6 well dish with Matrigel matrix for 1440 min or 1 hour.", "[Day 1] Thaw the frozen iPSCs by placing the vial in 37 °C water bath for 2 min.", "[Day 1] After thawing, spray the tube with 70% ethanol and place in biosafety cabinet.", "[Day 1] Aspirate the cells into 15 ml falcon tube and add 4 mL co...
19,997
U Mass - Aspartate Transferase
null
dx.doi.org/10.17504/protocols.io.xr5fm86
null
Jason Kim
TITLE: U Mass - Aspartate Transferase AUTHORS: Jason Kim [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span><span style = "font-weight:bold;"> </span></div><div class = "text-block">This experiment involves a spectrophotometric measurement using Roche Co...
["Perform daily quality control assessment of instrumentation before analysis.", "Load each sample into a specialized micro-sample cup for the clinical chemistry analyzer.", "Select Aspartate Transferase test on display and run the analysis.", "Collect and analyze the data."]
68,486
cDNA Library Preparation for scRNA-seq of Human Meniscus (10x Genomics)
5
dx.doi.org/10.17504/protocols.io.e6nvwkw32vmk/v1
https://www.protocols.io/view/cdna-library-preparation-for-scrna-seq-of-human-me-ce5etg3e
molmer, Martin Lotz, Tony Mondala, Steven Head
TITLE: cDNA Library Preparation for scRNA-seq of Human Meniscus (10x Genomics) AUTHORS: molmer, Martin Lotz, Tony Mondala, Steven Head [DESCRIPTION] Samples are processed using V2 barcoding chemistry kits of 10x Genomics. For each run, 10,000 cells from individual donors are labeled with distinct oligo-barcoded antib...
["Single cells are isolated from human knee menicus using doi dx.doi.org/10.17504/protocols.io.n2bvj6k7blk5/v1.", "Single cell suspensions are multiplexed and converted to barcoded scRNAseq libraries using the Chromium Single Cell 3’ Library, Gel Bead and Chip Kit (10x Genomics), and the BD™ Hu Single Cell Sample Multi...
null
null
null
dx.doi.org/10.17504/protocols.io.ssheeb6
null
null
TITLE: No Title AUTHORS: [BEFORE_START] <p>Clean the work area with 70% alcohol. Use all filters and autoclaved tips. Refrigerate your centrifuge to 4C.</p> [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.jqicmue
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
28,533
Preparing Reads for Stranded Mapping
null
dx.doi.org/10.17504/protocols.io.74vhqw6
null
David A. Eccles
TITLE: Preparing Reads for Stranded Mapping AUTHORS: David A. Eccles [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for preparing long reads for stranded mapping, as an intermediate step for additional protocols:</div><div class = "text-block"><ul style = "list-style-type:disc;"><l...
["[Index Preparation]\nPrepare transcript index (see Guidelines for data sources)lastdb Mus_musculus.GRCm38.cds.all.fa", "[Transcriptome Mapping]\nReads are mapped to the transcriptome with LAST.The results of that mapping can be piped through last-map-probs to exclude unlikely hits, then through 'maf-convert -n tab' t...
54,185
CoBG-11 preparation
4
dx.doi.org/10.17504/protocols.io.by6hpzb6
https://www.protocols.io/view/cobg-11-preparation-by6hpzb6
Arshshaikh
TITLE: CoBG-11 preparation AUTHORS: Arshshaikh [DESCRIPTION] CoBG-11 is a coculture medium used to coculture cyanobacteria and E. coli by Zhang et al (2020). It is optimized for E. coli growth. Here are its components: 1. 150 mM NaCl, 2. 4 mM NH4Cl 3. 3 g/L 2-[[1,3-dihydroxy-2-(hydroxymethyl) propan-2-yl] amino] et...
["Measure 0.88 g , 0.0214 g, and 0.3 g and add it to a 150mL flask.", "Add 50 mL to the flask and mix till contents dissolve. If salt persists, add 10mL more of BG-11 and mix well.", "Once contents dissolve, add BG-11 up to 95 mL.", "Adjust pH of the solution to 8.3 with NaOH.", "Fill BG-11 up to 100 mL measure pH and ...
32,343
Protocols for predicting Sphagnum bogs distributional pattern in China
null
dx.doi.org/10.17504/protocols.io.bbtxinpn
null
Mingyang Cong
TITLE: Protocols for predicting Sphagnum bogs distributional pattern in China AUTHORS: Mingyang Cong [STEPS] ?. Collect the center points of Sphagnum bogs. To collect the longitude and latitude for Sphagnum bogs’ central points across China, we principally consulted Swamps in China and Marshes in China and then extrac...
["Collect the center points of Sphagnum bogs. To collect the longitude and latitude for Sphagnum bogs’ central points across China, we principally consulted Swamps in China and Marshes in China and then extracted occurrence data from the literature. For the records lacking specific geographic coordinates, we used Googl...
93,523
JMN-MSMP NIA Automated Histological Staining
1
dx.doi.org/10.17504/protocols.io.eq2lyjb6rlx9/v1
https://www.protocols.io/view/jmn-msmp-nia-automated-histological-staining-c7jtzknn
ccherry
TITLE: JMN-MSMP NIA Automated Histological Staining AUTHORS: ccherry [DESCRIPTION] SenNet aged mouse histological staining from NIA [STEPS] SECTION: Fixation 1. Take the whole carcass into a jar and fix in formalin for 24 hr SECTION: Fixation 2. Wash the carcass with 70% EtOH and store in EtOH jar until histogrossing...
["[Fixation] Take the whole carcass into a jar and fix in formalin for 24 hr", "[Fixation] Wash the carcass with 70% EtOH and store in EtOH jar until histogrossing", "[Tissue collection:] Take out the ovaries together with uterus and vagina from the female samples and put into a histology cassette.", "[Tissue collectio...
81,475
Duragel Application for Acute Electrophysiology Recordings
1
dx.doi.org/10.17504/protocols.io.14egn2dwqg5d/v1
https://www.protocols.io/view/duragel-application-for-acute-electrophysiology-re-cttbwnin
anna.lakunina
TITLE: Duragel Application for Acute Electrophysiology Recordings AUTHORS: anna.lakunina [DESCRIPTION] This protocol describes the process of coating an implant with duragel in preparation for in vivo electrophysiology experiments. Duragel seals and protects the surface of the brain while also being soft enough for si...
["[Setup] Draw up the white and blue duragel components into separate 1 mL Luer lock syringes. Attach 20G needles.", "[Setup] Turn on microscope lights and heating pad.", "[Setup] Wrap heating pad in press ‘n’ seal.", "[Setup] Anesthetize the mouse (see section Prepare the anesthesia system and anesthetize the mouse (a...
39,605
SOLUTION- 08 - Türk solution
3
dx.doi.org/10.17504/protocols.io.biwvkfe6
https://www.protocols.io/view/solution-08-t-rk-solution-biwvkfe6
Marco Cosentino, Elisa Storelli, Alessandra Luini, Massimiliano Legnaro, Emanuela Rasini, Marco Ferrari, Franca Marino
TITLE: SOLUTION- 08 - Türk solution AUTHORS: Marco Cosentino, Elisa Storelli, Alessandra Luini, Massimiliano Legnaro, Emanuela Rasini, Marco Ferrari, Franca Marino [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This recepe is used in the following protocols:</div><div class = "text-block"><span sty...
[]
35,878
Triage for critically ill patients under the limited medical resource capacity during the COVID-19 and other infectious disease pandemics: a scoping review.
null
dx.doi.org/10.17504/protocols.io.bfaejibe
https://www.protocols.io/view/triage-for-critically-ill-patients-under-the-limit-bfaejibe
Yusuke Tsutsumi, Yasushi Tsujimoto, Sei Takahashi, Asuka Tsuchiya, Ikuyo Tsutsumi
TITLE: Triage for critically ill patients under the limited medical resource capacity during the COVID-19 and other infectious disease pandemics: a scoping review. AUTHORS: Yusuke Tsutsumi, Yasushi Tsujimoto, Sei Takahashi, Asuka Tsuchiya, Ikuyo Tsutsumi [DESCRIPTION] <div class = "text-blocks"><div class = "text-bloc...
[]
96,908
Mammalian cell culture and transfection for stable cell lines generation
0
dx.doi.org/10.17504/protocols.io.kxygxyk9dl8j/v1
https://www.protocols.io/view/mammalian-cell-culture-and-transfection-for-stable-davk2e4w
Hina Ojha, Miratul M. K. Muqit
TITLE: Mammalian cell culture and transfection for stable cell lines generation AUTHORS: Hina Ojha, Miratul M. K. Muqit [DESCRIPTION] Autosomal recessive mutations in PTEN-induced kinase 1 (PINK1) are linked to early-onset Parkinson's disease (PD) [1]. Upon mitochondrial depolarization, PINK1 activates through autopho...
["[Cell Culture] Maintain cells at 37 °C in a 5% CO2 water-saturated incubator.", "[Generation of Stable Cell Lines:] Achieve doxycycline-induced, stable expression of exogenous protein using the Flp-In T-Rex system according to Invitrogen's instructions, utilizing CRISPR knock-out PINK1 KO HeLa Flp-In T-Rex cells [4]....
88,334
High-throughput analysis of products from deconstructed nylon-6 by UHPLC-MS/MS (dMRM)
6
dx.doi.org/10.17504/protocols.io.6qpvr3k92vmk/v1
https://www.protocols.io/view/high-throughput-analysis-of-products-from-deconstr-c2hnyb5e
Kelsey J. Ramirez, Morgan A Ingraham, Elizabeth L. Bell, Gregg T. Beckham
TITLE: High-throughput analysis of products from deconstructed nylon-6 by UHPLC-MS/MS (dMRM) AUTHORS: Kelsey J. Ramirez, Morgan A Ingraham, Elizabeth L. Bell, Gregg T. Beckham [DESCRIPTION] A 3 minute, high-throughput analysis method was developed for the quantitation of products produced by deconstruction of nylon-6 ...
["[Preparation of Standards] By weight, create individual 2000 µg/mL stock solutions of all analytes listed below using ultrapure water (18.2MΩ⋅cm)(UPW) water as a diluent, except for 6-aminohexanoic acid cyclic-dimer in which methanol is used:\nɛ-Caprolactam\n6-Aminohexanoic acid\n6-Aminohexanoic acid dimer (6-(6-amin...
28,729
Nanopore Data Analysis
null
dx.doi.org/10.17504/protocols.io.8azhsf6
null
David A. Eccles
TITLE: Nanopore Data Analysis AUTHORS: David A. Eccles [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This is a collection of protocols that I use frequently for the analysis of long read data</div></div> [STEPS]
[]
44,236
General Salts + Sodium Bicarbonate Solutions
1
dx.doi.org/10.17504/protocols.io.bpfkmjkw
https://www.protocols.io/view/general-salts-sodium-bicarbonate-solutions-bpfkmjkw
Ada de la Cruz
TITLE: General Salts + Sodium Bicarbonate Solutions AUTHORS: Ada de la Cruz [STEPS] ?. [General Salts Solution:] For 1L of 10x salts solution:1 L nanopore water into 1 L bottle with lid55 g of MgCl2*6H2O5 g of NH4Cl1.4 g CaCl2*2H2O1 g KCl ?. [Sodium Bicarbonate] For 1 L of NaHCO3 (sodium bicarbonate):1 L nanopore wat...
["[General Salts Solution:]\nFor 1L of 10x salts solution:1 L nanopore water into 1 L bottle with lid55 g of MgCl2*6H2O5 g of NH4Cl1.4 g CaCl2*2H2O1 g KCl", "[Sodium Bicarbonate]\nFor 1 L of NaHCO3 (sodium bicarbonate):1 L nanopore water in a 1 L flask/bottle85 g of NaHCO3", "[General Salts Solution:]\nPour everything...
null
null
null
dx.doi.org/10.17504/protocols.io.cq5vy5
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?.
[]
45,442
HuBMAP Donor and Tissue Eligibility Criteria Form v 1.0
1
dx.doi.org/10.17504/protocols.io.bqmamu2e
https://www.protocols.io/view/hubmap-donor-and-tissue-eligibility-criteria-form-bqmamu2e
Yiing Lin, Shin Lin
TITLE: HuBMAP Donor and Tissue Eligibility Criteria Form v 1.0 AUTHORS: Yiing Lin, Shin Lin [STEPS] ?. HuBMAP Donor and Tissue Eligibility Criteria FormVersion 1.0Effective Date: 1/1/2019 Inel...
["HuBMAP Donor and Tissue Eligibility Criteria FormVersion 1.0Effective Date: 1/1/2019 Ineligible EligibleOverall eligibilityDoes the consent allow for research activities? ...
43,671
pYCR cloning strategy
1
dx.doi.org/10.17504/protocols.io.bnvxme7n
https://www.protocols.io/view/pycr-cloning-strategy-bnvxme7n
Andreea S
TITLE: pYCR cloning strategy AUTHORS: Andreea S [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Introducing NLP14a in the genome of B. mycoides: </span><span> In order to create plasmid pYCR-gamyNLP, the backbone of Pycr will be digested with the PCR product of gam...
["[Clone sgRNA sequence into pYCR]\nDesign sgRNA spacer sequence (~20 nts) using \"Benchling'\" and choose B. mycoides M2E_15 genomeChoose the gRNA with the highest on-target score and the lowest off-target score.", "[Clone sgRNA sequence into pYCR]\nOrder the 2 complementary oligos flanked by overhang containing BsaI ...
54,234
Cecret Workflow for SARS-CoV-2 Assembly and Lineage Classification
5
null
https://www.protocols.io/view/cecret-workflow-for-sars-cov-2-assembly-and-lineag-by72pzqe
Erin L Young, Technical Outreach and Assistance for States Team
TITLE: Cecret Workflow for SARS-CoV-2 Assembly and Lineage Classification AUTHORS: Erin L Young, Technical Outreach and Assistance for States Team [DESCRIPTION] This protocol provides instructions to install and run the Cecret workflow as part of the StaPH-B Toolkit. Cecret produces SARS-CoV-2 consensus sequence ass...
["[Software Dependencies] Load software dependencies\n \n\nWithin certain high-performance computing environments, these software can be loaded using GNU module commands similar to:", "[Installing StaPH-B Toolkit] The Cecret assembly workflow can be installed as part of the StaPH-B Toolkit using the following commands:...
62,875
Prima UK--Reviews-2022
1
dx.doi.org/10.17504/protocols.io.14egn7nzqv5d/v1
https://www.protocols.io/view/prima-uk-reviews-2022-b9m3r48n
prima
TITLE: Prima UK--Reviews-2022 AUTHORS: prima [DESCRIPTION] Prima UK--Reviews-2022 [STEPS] SECTION: How does Prima UK Function? 1. hat's going on here? The Prima Keto UK or "keto" diet is a low-carb, fat-rich eating plan that has been utilized for a really long time to treat explicit ailments. In the nineteenth 100 ...
["[How does Prima UK Function?] hat's going on here?\nThe Prima Keto UK or \"keto\" diet is a low-carb, fat-rich eating plan that has been utilized for a really long time to treat explicit ailments. In the nineteenth 100 years, the Prima Keto UK was generally used to assist with controlling diabetes. In 1920 it was pre...
87,626
Sanger Tree of Life Sample Homogenisation: Cryogenic Bead Beating of Plants with FastPrep-96
4
dx.doi.org/10.17504/protocols.io.rm7vzxk38gx1/v1
https://www.protocols.io/view/sanger-tree-of-life-sample-homogenisation-cryogeni-cztix6ke
Benjamin Jackson, Caroline Howard
TITLE: Sanger Tree of Life Sample Homogenisation: Cryogenic Bead Beating of Plants with FastPrep-96 AUTHORS: Benjamin Jackson, Caroline Howard [DESCRIPTION] Disruption under cryogenic conditions is vital to interrupt the often tough, flexible and rigid structure of plant tissues whilst maintaining nucleic acid integri...
["[Laboratory protocol] Prepare all necessary equipment prior to starting and place any applicable items (e.g. cold blocks, tools) onto dry ice.", "[Laboratory protocol] Add 3 x 3 mm sterile stainless steel beads (Qiagen PN 69997) to the required number of 1.9 mL Tri-coded FluidX tubes and place into a cold block on dr...
null
null
null
dx.doi.org/10.17504/protocols.io.ke6cthe
null
null
TITLE: No Title AUTHORS: [STEPS]
[]
85,705
Protocol for Facially Guided Digital Diagnosis in Orthodontics
1
dx.doi.org/10.17504/protocols.io.8epv5x9q6g1b/v1
https://www.protocols.io/view/protocol-for-facially-guided-digital-diagnosis-in-cxxhxpj6
Rupert HG Kelley BSc, Álvaro Ferrando Cascales DDS,PhD, Raúl Ferrando Cascales DDS,MSc,PhD
TITLE: Protocol for Facially Guided Digital Diagnosis in Orthodontics AUTHORS: Rupert HG Kelley BSc, Álvaro Ferrando Cascales DDS,PhD, Raúl Ferrando Cascales DDS,MSc,PhD [DESCRIPTION] As the digital age of dentistry continues to flourish, it has never been more important to have protocols to guide dentists through the...
["[Treatment Planning] Records: NHP pictures, STL and DICOM files", "[Treatment Planning] Orientation and alignment of STL and DICOM files with photographs\n\nOnce all the records have been obtained, the digital impressions (STLs) can be superimposed onto the natural head position photographs using a software package s...
41,232
allele.variability
1
dx.doi.org/10.17504/protocols.io.bkhqkt5w
https://www.protocols.io/view/allele-variability-bkhqkt5w
Sara Beier, Sara Beier
TITLE: allele.variability AUTHORS: Sara Beier, Sara Beier [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">A prerequisite to improve the predictability of microbial community dynamics is to understand their assembly mechanisms. To study factors that contribute to microbial community assembly, we exam...
["removal of nextera adaptors (cutadapt v1.8.3)"]
70,911
Planktoscope protocol for plankton imaging
1
dx.doi.org/10.17504/protocols.io.bp2l6bq3zgqe/v2
https://www.protocols.io/view/planktoscope-protocol-for-plankton-imaging-chg7t3zn
Lombard Fabien, Will Major, Anna Oddone, Clémence Clausse
TITLE: Planktoscope protocol for plankton imaging AUTHORS: Lombard Fabien, Will Major, Anna Oddone, Clémence Clausse [DESCRIPTION] this protocol is for using planktoscope and collect usable result for quantitative imaging of plankton see also https://www.planktoscope.org/ [BEFORE_START] -Test the protocol before acq...
["[Calibration] Pump calibration:", "[Calibration] -prepare a large volume of tap water and put in in the syringe targeting a total volume of e.g. 20ml\n-on the optic configuration tab: tell him that you want to pass 10ml and record the exact volume it finally ends to pass (eg. by looking on the graduation of the syrin...
12,419
AMPureXP purification
1
dx.doi.org/10.17504/protocols.io.4r3l2wy3l1y9/v1
https://www.protocols.io/view/ampurexp-purification-qdbds2n
Eva Petrova, Roey Angel
TITLE: AMPureXP purification AUTHORS: Eva Petrova, Roey Angel [DESCRIPTION] PCR-product purification with AMPureXP solution. From the manual: "The Agencourt AMPure XP system is a highly efficient, easily automated PCR purification system that delivers superior-quality DNA with no salt carryover. Requiring no centrif...
["Shake the Agencourt AMPureXP bottle to fully resuspend magnetic particles.", "Add sample Vol µL × 1.8 (or less i.e. 0.8) of the Agencourt AMPure XP solution. Pipette mix 10 times, or vortex a few seconds.", "Incubate at room temperature for 5 minutes.\n5 min", "Place the reaction plate/tube onto an Agencourt SPRIPlat...
60,289
2022 GenomeTrakr Proficiency Testing exercise (PulseNet Harmonized)
1
dx.doi.org/10.17504/protocols.io.e6nvw5m9dvmk/v4
https://www.protocols.io/view/2022-genometrakr-proficiency-testing-exercise-puls-b649rgz6
Maria Balkey, Ruth Timme, Julie Haendiges
TITLE: 2022 GenomeTrakr Proficiency Testing exercise (PulseNet Harmonized) AUTHORS: Maria Balkey, Ruth Timme, Julie Haendiges [DESCRIPTION] This SOP outlines guidelines on how to process the isolates for the 2022 GenomeTrakr (GT) Proficiency Testing exercise. This SOP is applicable to all GenomeTrakr labs partici...
["[Culture Preparation] Salmonella and Escherichia/Shigella Lyophilized cultures:\n\nDay 1\n\nDocument the isolate number(s) and the lyophilized date(s) for your records. Wipe the aluminum cover and outside of the vial with isopropyl alcohol. Using sturdy forceps, aseptically remove the aluminum cover and rubber stoppe...
99,199
Protocol (B): Zebrafish embedding and imaging (3 dpf)
0
dx.doi.org/10.17504/protocols.io.14egn6726l5d/v1
https://www.protocols.io/view/protocol-b-zebrafish-embedding-and-imaging-3-dpf-dc472yzn
Désirée A. Schmitz, Tobias Wechsler, Hongwei Bran Li, Bjoern H. Menze, Rolf Kümmerli
TITLE: Protocol (B): Zebrafish embedding and imaging (3 dpf) AUTHORS: Désirée A. Schmitz, Tobias Wechsler, Hongwei Bran Li, Bjoern H. Menze, Rolf Kümmerli [DESCRIPTION] This protocol details the zebrafish embedding and imaging. [STEPS] SECTION: Part 0: Material preparation 1. Prepare 1.5% low-melting point agarose by...
["[Part 0: Material preparation] Prepare 1.5% low-melting point agarose by heating e.g. 1.5 g low-melting point agarose (Sigma, serial no: A9414) in 100 mL distilled water in a flask (minimal volume 150 mL) in 30 s bursts in the microwave.", "[Part 0: Material preparation] Once the solution is clear, i.e. no flocs or p...
28,839
Microfluidic Digital Logic Chip Assembly
1
dx.doi.org/10.17504/protocols.io.8efhtbn
https://www.protocols.io/view/microfluidic-digital-logic-chip-assembly-8efhtbn
Erik Werner, Elliot Hui [University Of California
TITLE: Microfluidic Digital Logic Chip Assembly AUTHORS: Erik Werner, Elliot Hui [University Of California [DESCRIPTION] This protocol describes how to reversibly bond layers of poly(methyl methacrylate) (PMMA) and polydimethylsiloxane (PDMS) using thermo-compresison bonding. Usually, two layers of PMMA are sandwiched...
["[Clean] Rinse all parts to be assembled under a stream of DI water to remove charred material or debris from machining.", "[Clean] Prepare a solution of ~1% Microsoap in DI water and fill each bag about half way", "[Clean] Insert each component of the Microfluidic device into a 3\" x 4\" zip loc bag.", "[Clean] Sonic...
52,679
Investigation_of_mitophagy_in_Hippo_neurons
1
dx.doi.org/10.17504/protocols.io.bxpfpmjn
https://www.protocols.io/view/investigation-of-mitophagy-in-hippo-neurons-bxpfpmjn
OLIVIA HARDING, Chantell S. Evans
TITLE: Investigation_of_mitophagy_in_Hippo_neurons AUTHORS: OLIVIA HARDING, Chantell S. Evans [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">We developed a method for assessing mitochondrial clearance in primary hippocampal neurons. </div></div> [STEPS] ?. [Plating and maintenance of hippocampal ...
["[Plating and maintenance of hippocampal neurons]\nPlate primary hippocampal Sprague Dawley rat neurons (embryonic day 18) on 35 mm glass bottomed dishes at 250,00 cells per dish in MEM supplemented with 10% horse serum, 33 mM D-glucose, and 1 mM sodium pyruvate.", "[Plating and maintenance of hippocampal neurons]\nAd...
25,252
Loop L2 (even level) type IIS cloning - pCs-ye vectors
null
dx.doi.org/10.17504/protocols.io.4wcgxaw
null
Eftychis Frangedakis, Susana Sauret-Gueto, Anthony West, marta tomaselli, Nicola Patron, Marius Rebmann, Jim Haseloff
TITLE: Loop L2 (even level) type IIS cloning - pCs-ye vectors AUTHORS: Eftychis Frangedakis, Susana Sauret-Gueto, Anthony West, marta tomaselli, Nicola Patron, Marius Rebmann, Jim Haseloff [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol based on</div><div class = "text-block"><a href="http:...
["Determine DNA parts concentration with spectrophotometry (Nanodrop).", "Prepare aliquots for DNA parts to be assembled at a concentration of 15 nM and of the pCs-pe vector at a concentration of 7.5 nM.To calculate the concentration needed for each part to assemble (not the backbone) in ng/µL, divide the length of the...
78,527
A Systematic Review of Bullous Pemphigoid and HLA-DQA1
1
dx.doi.org/10.17504/protocols.io.14egn2r6pg5d/v1
https://www.protocols.io/view/a-systematic-review-of-bullous-pemphigoid-and-hla-cqw7vxhn
Dylan Thibaut, Roksana Hesari, Nina Schur, Shivani Thoutireddy, Ryan Witcher, Elyse Julian
TITLE: A Systematic Review of Bullous Pemphigoid and HLA-DQA1 AUTHORS: Dylan Thibaut, Roksana Hesari, Nina Schur, Shivani Thoutireddy, Ryan Witcher, Elyse Julian [DESCRIPTION] There is growing evidence suggesting that specific HLA-DQA1 alleles are associated with increased odds of developing bullous pemphigoid. Howeve...
["[Administrative Information] Tilte:\n\n\"A Systematic Review of Bullous Pemphigoid and HLA-DQA1\"\n\nRegistrations:\n\nprotocols.io", "[Methods] Eligibility Criteria \n\nInclusion Criteria:\n1. Studies that investigate the association between HLA-DQA1 and bullous pemphigoid in human subjects.\n2. Studies published af...
null
null
null
dx.doi.org/10.17504/protocols.io.s4regv6
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Transformation of <em>Acanthamoeba castellanii </em>Neff (ATCC-30010) with plasmid DNA can be achieved with a modified version of the protocol described in Peng, Omaruddin, &amp; Bateman (2005) using constructs from Bateman (2010). </p> [BEFORE_START] <p>Materials needed:</p...
[]
50,824
Nissl Staining
1
dx.doi.org/10.17504/protocols.io.bp2l6bz9zgqe/v1
https://www.protocols.io/view/nissl-staining-bvvgn63w
Pranay Srivastava, Xiqun Chen
TITLE: Nissl Staining AUTHORS: Pranay Srivastava, Xiqun Chen [DESCRIPTION] This protocol details the staining procedure and mounting procedure of Nissl staining. [STEPS] SECTION: Staining Procedure 1. Mount sections on positive charged plus slides. Air dry sections . SECTION: Staining Procedure 2. Incubate the slid...
["[Staining Procedure] Mount sections on positive charged plus slides. Air dry sections .", "[Staining Procedure] Incubate the slides directly into 1:1 ethanol/chloroform for 30 min.\n(All the steps mentioned should be done under the hood)", "[Staining Procedure] Incubate in 100% ethanol 2 min", "[Staining Procedure]...
86,287
W-2 WATER PROCESSING
4
dx.doi.org/10.17504/protocols.io.ewov1opwplr2/v1
https://www.protocols.io/view/w-2-water-processing-cyhpxt5n
REDI-NET Consortium
TITLE: W-2 WATER PROCESSING AUTHORS: REDI-NET Consortium [DESCRIPTION] This protocol details about water processing. [BEFORE_START] BEFORE START Make sure the feeding tube and PTFE sinker for SolVac Filter are properly clean by using 70% ethanol and allowed to air dry. For the samples stored at 4 °C, pour thre...
["[1. VACUUM PUMP SET UP] Wipe the surfaces with 70% ethanol to remove contaminants.", "[1. VACUUM PUMP SET UP] Use tubing to connect a 3 liter Medi-Vac Canister with vacuum pump through the vacuum outlet on the lid.", "[1. VACUUM PUMP SET UP] Connect tubing with the 3 liter Medi-Vac Canister through the inlet on the l...
85,912
Typology of Physical Activity
1
dx.doi.org/10.17504/protocols.io.yxmvm32kbl3p/v1
https://www.protocols.io/view/typology-of-physical-activity-cx5yxq7w
Christine Roberts
TITLE: Typology of Physical Activity AUTHORS: Christine Roberts [DESCRIPTION] Different physical activity types vary in metabolic demand (intensity), but also in non-metabolic demand (balance, co-ordination, speed and flexibility), cognitive demand (attention, memory and decision making), and social demand (social int...
["[Typology of Physical Activity] A TYPOLOGY OF PHYSICAL ACTIVITY", "[Typology of Physical Activity] Title: A Typology of Physical Activity", "[Typology of Physical Activity] Investigators: Chief Investigator: Christine Roberts, University of Aberdeen, Health Psychology, Health Sciences Building, Foresterhill, Aberdeen...
27,805
SMARTerV4 (0.5x) Amplification for single-cell or single-nuclei RNASeq Protocol
null
dx.doi.org/10.17504/protocols.io.7d5hi86
null
ZengU19 BRAIN grant
TITLE: SMARTerV4 (0.5x) Amplification for single-cell or single-nuclei RNASeq Protocol AUTHORS: ZengU19 BRAIN grant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol to generate full-length cDNA from single cells, or nuclei, using Takara SMARTer V4.</div></div> [STEPS]
[]
104,146
Developing and Testing Framework for Assessing Quality of Facility Readiness for Clinical Teaching and Learning in Tanzania - A Mixed Method Study.
0
dx.doi.org/10.17504/protocols.io.x54v92kn1l3e/v1
https://www.protocols.io/view/developing-and-testing-framework-for-assessing-qua-dhxs37ne
Jackson Karani Maira
TITLE: Developing and Testing Framework for Assessing Quality of Facility Readiness for Clinical Teaching and Learning in Tanzania - A Mixed Method Study. AUTHORS: Jackson Karani Maira [DESCRIPTION] Background Clinical education and clinical learning are important parts of nursing education, providing students with th...
["[Developing and Testing Framework for Assessing Quality of Facility Readiness for Clinical Teaching and Learning in Tanzania - A Mixed Method Study.] ABSTRACT\nBackground\nClinical education and clinical learning are important parts of nursing education,\nproviding students with the hands-on experience and practical ...