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97,191
Plate Scale Tn5 based tagmentation library prep protocol
1
dx.doi.org/10.17504/protocols.io.4r3l2qmzpl1y/v1
https://www.protocols.io/view/plate-scale-tn5-based-tagmentation-library-prep-pr-da6f2hbn
Cade Mirchandani, max genetti, Pingting Wang, Evan Pepper-Tunick, Shelbi Russell, Russell Corbett-Detig
TITLE: Plate Scale Tn5 based tagmentation library prep protocol AUTHORS: Cade Mirchandani, max genetti, Pingting Wang, Evan Pepper-Tunick, Shelbi Russell, Russell Corbett-Detig [DESCRIPTION] This protocol is a fork of (dx.doi.org/10.17504/protocols.io.bv5gn83w), which a demonstrated efficient and high-throughput tagme...
["[Transposon Assembly] Remove oligos Tn5ME-A, Tn5ME-B, Tn5ME-R, and Tn5 enzyme mix from freezer. Thaw primers, mix by vortexing and spin. Keep Tn5 enzyme on ice. Turn on 95 °C thermocycler.", "[Transposon Assembly] In separate PCR tubes combine:\n7 µL Tn5ME-A + 7 µL Tn5ME-R = AR oligo (14 µL)\n7 µL Tn5ME-B + 7 µL Tn5M...
28,601
Disruption of Synechocystis cells
null
dx.doi.org/10.17504/protocols.io.76zhrf6
null
iGEM Dusseldorf
TITLE: Disruption of Synechocystis cells AUTHORS: iGEM Dusseldorf [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for disrupting Synechocystis cells to get soluble and non-soluble protein in different fractions</div><div class = "text-block"><ul style = "list-style-type:disc;"><li style = "...
[]
25,834
Calibration Protocol - OD600 Inter-equipment Conversion with LUDOX
null
dx.doi.org/10.17504/protocols.io.5gig3ue
null
Paul Rutten, Richard Tennant, Jacob Beal, Traci Haddock-Angelli, Natalie Farny, Geoffrey Baldwin, Marko Storch, Ari Dwijayanti
TITLE: Calibration Protocol - OD600 Inter-equipment Conversion with LUDOX AUTHORS: Paul Rutten, Richard Tennant, Jacob Beal, Traci Haddock-Angelli, Natalie Farny, Geoffrey Baldwin, Marko Storch, Ari Dwijayanti [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align...
["[Data collection: OD 600 Reference point - LUDOX Protocol]\nAdd 100 μl LUDOX CL-X into wells A1, B1, C1, D1", "[Data collection: OD 600 Reference point - LUDOX Protocol]\nAdd 100 μl of ddH20 into wells A2, B2, C2, D2", "[Data collection: OD 600 Reference point - LUDOX Protocol]\nMeasure absorbance at 600 nm of all sa...
81,462
10x Protocols: Visium v2 CytAssist FFPE Library Construction -- University of Minnesota TMCs (CG000495 Rev C)
1
dx.doi.org/10.17504/protocols.io.e6nvwj3zdlmk/v1
https://www.protocols.io/view/10x-protocols-visium-v2-cytassist-ffpe-library-con-ctswwnfe
IOx Genomics, Laura J Niedernhofer, David A Bernlohr
TITLE: 10x Protocols: Visium v2 CytAssist FFPE Library Construction -- University of Minnesota TMCs (CG000495 Rev C) AUTHORS: IOx Genomics, Laura J Niedernhofer, David A Bernlohr [DESCRIPTION] Protocols from 10x Genomics for Visium Spatial Gene Expression v2 chemistry on FPPE samples with the CytAssist component. Pro...
["10x protocol CG000495, Revision C (Library construction with CytAssist)", "Additional Protocols/Resources\nhttps://www.10xgenomics.com/support/spatial-gene-expression-ffpe"]
null
null
null
dx.doi.org/10.17504/protocols.io.cqavsd
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?.
[]
47,622
Expression and purification of Tribolium castaneum orthologue of PINK1
1
dx.doi.org/10.17504/protocols.io.bsrend3e
https://www.protocols.io/view/expression-and-purification-of-tribolium-castaneum-bsrend3e
Olawale G. Raimi, Miratul M. K. Muqit
TITLE: Expression and purification of Tribolium castaneum orthologue of PINK1 AUTHORS: Olawale G. Raimi, Miratul M. K. Muqit [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Mutations in PINK1 (protein kinase) and Parkin (ubiquitin E3 ligase) have been linked to familial early-onset Parkinson’s disea...
["[Transformation of competent bacteria]\nMix of pET 6His sumo codon optimised TcPINK1 150-end ΔI261-L270 S205E E527A K528A plasmid () with of the competent BL21(DE3) cells.\n1 µl\n50 µl", "[Transformation of competent bacteria]\nIncubate for .\non ice", "[Transformation of competent bacteria]\nHeat shock the cells i...
92,308
Protocol for performing PINK1 siRNA knockdown in mouse embryonic fibroblasts (MEFs)
1
dx.doi.org/10.17504/protocols.io.kxygx343zg8j/v1
https://www.protocols.io/view/protocol-for-performing-pink1-sirna-knockdown-in-m-c6duza6w
Enrico Bagnoli, Miratul Muqit
TITLE: Protocol for performing PINK1 siRNA knockdown in mouse embryonic fibroblasts (MEFs) AUTHORS: Enrico Bagnoli, Miratul Muqit [DESCRIPTION] This protocol details the siRNA knockdown in mouse embryonic fibroblasts (MEFs) for PINK1, but applicable for any other target. [GUIDELINES] 72-Hr siRNA knockdown in MEFs in ...
["[Day 1 - Cell Seeding and siRNA preparation] For Primary MEFs seed 200,000 cells per well in a 6-well plate at a total volume of 2 mL.", "[Day 1 - Cell Seeding and siRNA preparation] Add 400 µL of RNA-free water.", "[Day 1 - Cell Seeding and siRNA preparation] Incubate in hood for 5 min, vortex vigorously and store a...
71,512
Total RNA and DNA from Microalgae (12 samples per microplate)
1
dx.doi.org/10.17504/protocols.io.6qpvro85bvmk/v11
https://www.protocols.io/view/total-rna-and-dna-from-microalgae-12-samples-per-m-ch3yt8pw
Ying-Yu Hu, Zoe V. Finkel
TITLE: Total RNA and DNA from Microalgae (12 samples per microplate) AUTHORS: Ying-Yu Hu, Zoe V. Finkel [DESCRIPTION] Here we describe a protocol for extracting and quantifying bulk RNA and DNA from microalgae, which is adapted from Berdalet E. et al. (2005). RNA and DNA are extracted from microalgae samples and the...
["[Day 1: Freeze-dry samples] Freeze dry samples and blank filters. Freeze at -80 °C until processed.", "[Day 1: Prepare primary solutions] Turn on UV light in biosafety cabinet for 15 min", "[Day 1: Prepare primary solutions] Prepare Tris buffer 5 mM pH 8.0", "[Day 1: Prepare primary solutions] Pour 1 M pH 8.0 Tris ...
70,629
SOP for Spatial N-glycomics
1
null
https://www.protocols.io/view/sop-for-spatial-n-glycomics-cg8dtzs6
Chris Anderton
TITLE: SOP for Spatial N-glycomics AUTHORS: 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. [STEPS] SECTION: Scope 1. This protocol describes the procedure to obtain high quality ...
["[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.", "[Procedure] Formalin fixed tissue should...
106,703
Miniscope calcium imaging data acquisition of cortical activity in non-human primates (NHPs)
0
dx.doi.org/10.17504/protocols.io.14egn612pl5d/v1
https://www.protocols.io/view/miniscope-calcium-imaging-data-acquisition-of-cort-dkfp4tmn
Adriana Galvan, Thomas Wichmann
TITLE: Miniscope calcium imaging data acquisition of cortical activity in non-human primates (NHPs) AUTHORS: Adriana Galvan, Thomas Wichmann [DESCRIPTION] The protocol describes head-mounted miniscope calcium imaging data acquisition for NHPs. [GUIDELINES] Introduction: In recent years, calcium imaging has become a ...
["[Procedure] The ability to image calcium transients requires prior placement of a GRIN lens assembly (as detailed in a separate protocol dx.doi.org/10.17504/protocols.io.e6nvw15w2lmk/v1).", "[Procedure] Calcium imaging starts several weeks after the cortical implantation of the GRIN lens assembly and injection of a v...
28,216
Making and running an acrylamide protein gel
null
dx.doi.org/10.17504/protocols.io.7syhnfw
null
Robert Hooftman
TITLE: Making and running an acrylamide protein gel AUTHORS: Robert Hooftman [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">In this protocol, the preparation and running an acrylamide gel is explained. Using this gel, proteins with a size of >15 kDa can be visualized. For the visualization of prote...
["Make the separation gel. In the separation gel, the proteins are separated by size. The following is needed for 10 ml of separation gel (add in the following order): ABC1Ingredients10% acrylamide gel12% acrylamide gel2dH2O 4.1 ml 3.4 ml 3Acrylamide (30%, 37.5:1; Bio-Rad) 3.3 ml 4 ml 4Tris-HCl (1.5 M, pH 8.8) 2.5 ml ...
19,440
Mindfulness and Empathy University of Burgos
null
dx.doi.org/10.17504/protocols.io.w8qfhvw
null
Juan Pablo Pizarro, Raquel De la Fuente Anuncibay, Ángela González Barbadillo, González-Bernal, Esther Cubo
TITLE: Mindfulness and Empathy University of Burgos AUTHORS: Juan Pablo Pizarro, Raquel De la Fuente Anuncibay, Ángela González Barbadillo, González-Bernal, Esther Cubo [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Mediating effect of mindfulness cognition on the development of empathy in a univer...
[]
94,793
Wheat disease symptoms observation, capture, description, and evaluation
4
dx.doi.org/10.17504/protocols.io.ewov1q67kgr2/v1
https://www.protocols.io/view/wheat-disease-symptoms-observation-capture-descrip-c8thzwj6
Erin H Hill, Benjamin Schwessinger
TITLE: Wheat disease symptoms observation, capture, description, and evaluation AUTHORS: Erin H Hill, Benjamin Schwessinger [DESCRIPTION] This protocols is part of the ANU Biosecurity mini-research project #1 "Plant Pathogen Diagnostics: Visuals, subcultures, and genomics". You will be provided four pots of 3-4 week...
["[Disease symptom capture] You will receive five pots with wheat plants as detailed in the \"Description\" section. Please label each pot with your research group name, the date, and the treatment group.", "[Disease symptom capture] Carefully, study the plants in each pot. Select four to five leaves for each treatment...
62,878
Protocol: MHC class I and dengue hemorrhagic fever: a systematic review of HLA-A*24 and HLA-B*44
4
dx.doi.org/10.17504/protocols.io.b9m6r49e
https://www.protocols.io/view/protocol-mhc-class-i-and-dengue-hemorrhagic-fever-b9m6r49e
Andrew C Cook, Dylan Thibaut
TITLE: Protocol: MHC class I and dengue hemorrhagic fever: a systematic review of HLA-A*24 and HLA-B*44 AUTHORS: Andrew C Cook, Dylan Thibaut [DESCRIPTION] This project will examine the effect of two MHC class I alleles, HLA-A*24 and HLA-B*44 on dengue hemorrhagic fever susceptibility. A systematic review will be con...
["[Title] MHC class I and dengue hemorrhagic fever: a systematic review of HLA-A*24 and HLA-B*44", "[Introduction]", "[Methods]", "[Introduction] The association of HLA alleles with dengue fever and dengue hemorrhagic fever is a topic of research interest. Dengue fever has been evaluated for associations with HLA allel...
null
null
null
dx.doi.org/10.17504/protocols.io.rtfd6jn
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?. ?. ?. ?. ?.
["For inhibitors, freshly prepared aldose reductase (polyol pathway) inhibitor (Epalrestat, Sigma SML0527) and carnitine palmitoyltransferase-1 inhibitor (Etomoxir, Sigma E1905) were solubilised in water to prepare a 5 mM stock.", "Glycolysis inhibitor (2-Deoxy-D-Glucose, Sigma D8375) was solubilised in water to prepar...
90,729
Confocal-based bead-binding
4
dx.doi.org/10.17504/protocols.io.8epv5xdbng1b/v1
https://www.protocols.io/view/confocal-based-bead-binding-c4uhywt6
Dan Tudorica
TITLE: Confocal-based bead-binding AUTHORS: Dan Tudorica [DESCRIPTION] Method of quantifying in vitro binding. [STEPS] SECTION: Protein preparation 1. In order to prepare proteins for a fluorescent bead-binding experiment, you need one protein that can bind a resin (in my case MBP-Rubicon RH domain, which can bind ...
["[Protein preparation] In order to prepare proteins for a fluorescent bead-binding experiment, you need one protein that can bind a resin (in my case MBP-Rubicon RH domain, which can bind amylose resin) and a fluorescently labelled bait protein (in my case, Rab7-AlexaFluor 647).\n\nTo prepare my AlexaFluor 647, used a...
8,041
Carbonic Anhydrase Activity Assay
null
dx.doi.org/10.17504/protocols.io.j4hcqt6
null
Jack Koch, Virginia Weis
TITLE: Carbonic Anhydrase Activity Assay AUTHORS: Jack Koch, Virginia Weis [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span>Carbonic anhydrase (CA) is an enzyme which catalyzes the interconversion of bicarbonate ions (HCO</span><span style = "vertical-align:sub;vertical-align:super;">3</span><s...
["[Mixing Carbonic Anhydrase (CA) Buffers]\nStock 50mM barbital buffer", "[Mixing Carbonic Anhydrase (CA) Buffers]\n25 mM Veronal buffer (Buffer A)Mix 250mL of 50mM barbital buffer with 250mL of deionized waterAdjust pH of Buffer A to 8.2 with 1M HCl", "[Mixing Carbonic Anhydrase (CA) Buffers]\nExtraction bufferMix 250...
18,667
Cleaning up a biohazardous spill outside of a biosafety cabinet
null
dx.doi.org/10.17504/protocols.io.wgjfbun
null
Steven Wilhelm, Gary LeCleir, Ashley Humphrey
TITLE: Cleaning up a biohazardous spill outside of a biosafety cabinet AUTHORS: Steven Wilhelm, Gary LeCleir, Ashley Humphrey [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">BIOLOGICAL SPILL RESPONSE </span><span>method to be used in the event of a biosafety inciden...
["Close off the area and allow aerosols to settle.", "Notify others including supervisor.", "Assemble all spill cleanup materials and review procedure.", "Don appropriate PPE: laboratory coat, safety glasses or chemical splash goggles (depending on risk of splashes), Nitrile gloves.", "Cover spill with paper towels.", ...
40,289
Purification of anti-anti-SpA antibody (Ab-2) using SpA-bearing Staphylococcus aureus cells.
6
dx.doi.org/10.17504/protocols.io.bjj9kkr6
https://www.protocols.io/view/purification-of-anti-anti-spa-antibody-ab-2-using-bjj9kkr6
Angel Justiz-Vaillant
TITLE: Purification of anti-anti-SpA antibody (Ab-2) using SpA-bearing Staphylococcus aureus cells. AUTHORS: Angel Justiz-Vaillant [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">A novel method was designed for the purification of Ab-2 from undiluted WSF of hyperim...
["Mix in an Eppendorf microtube 0.9 ml of undiluted WSF from immunized birds with 25 μl of SpA-bearing Staphylococcus aureus cells (Sigma-Aldrich).", "Incubate the microtube at 37oC for 30 min.", "After the incubation period, centrifuge the microtube in an Eppendorf 5424 centrifuge for 5 min.", "Observe the microtube t...
null
null
null
dx.doi.org/10.17504/protocols.io.jk7ckzn
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol can be used for mapping transcriptional start site(s) (TSS) of a specific gene of interest in bacteria.</p> <p>By slightly modifying this protocol, whole transcriptome TSS could also potentially be mapped.</p> [BEFORE_START] <p>Oligos used:</p> <table style="wi...
[]
58,494
Pharmacokinetics and bioequivalence of two imidocarb formulations in cattle after subcutaneous injection
1
dx.doi.org/10.17504/protocols.io.b5c6q2ze
https://www.protocols.io/view/pharmacokinetics-and-bioequivalence-of-two-imidoca-b5c6q2ze
Anonymous , Chen Chen, Maolin Liu, Xiaojie Chen, Chunshuang Liu, Yanyan Feng, Xinbo Yan, Yiming Liu, Xiubo Li, Honglei Wang
TITLE: Pharmacokinetics and bioequivalence of two imidocarb formulations in cattle after subcutaneous injection AUTHORS: Anonymous , Chen Chen, Maolin Liu, Xiaojie Chen, Chunshuang Liu, Yanyan Feng, Xinbo Yan, Yiming Liu, Xiubo Li, Honglei Wang [DESCRIPTION] Imidocarb (IMD) is commonly used for treatment of eperythro...
["Sample collection \nBlood samples (10–15 mL) were collected from the jugular vein at 0 h before administration and 10 min, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h after subcutaneous administration, and were drawn in vacutainers containing disodium EDTA as anticoagulant. The samples were immediately centrif...
74,738
Recombinant protein expression and purification of fuGFP
4
dx.doi.org/10.17504/protocols.io.e6nvwje79lmk/v1
https://www.protocols.io/view/recombinant-protein-expression-and-purification-of-ck8suzwe
Javiera A Avilés, Tamara Matute, Isaac Núñez, Maira Rivera, Javiera Reyes, Anibal Arce Medina, Cesar A Ramirez-Sarmiento, Fernan Federici
TITLE: Recombinant protein expression and purification of fuGFP AUTHORS: Javiera A Avilés, Tamara Matute, Isaac Núñez, Maira Rivera, Javiera Reyes, Anibal Arce Medina, Cesar A Ramirez-Sarmiento, Fernan Federici [DESCRIPTION] This protocol has been optimized for the recombinant expression of fuGFP encoded in an open ...
["[DAY 1 – Plasmid transformation] Transform 100 ngof the open pTi plasmid containing fuGFP into E. coli BL21 (DE3) competent cells using either heat shock or electroporation.", "[DAY 1 – Plasmid transformation] Spread transformed cells in LB Agar plates supplemented with 0.05 mg/mLKan. Grow plate overnight at 37 °C.",...
58,696
Biolistic transformation of Pseudo-nitzschia multistriata
4
dx.doi.org/10.17504/protocols.io.b5jgq4jw
https://www.protocols.io/view/biolistic-transformation-of-pseudo-nitzschia-multi-b5jgq4jw
Anna Santin, Monia Teresa Russo, Pina Marotta, Francesco Manfellotto, Antonella Ruggiero, Mariella Ferrante
TITLE: Biolistic transformation of Pseudo-nitzschia multistriata AUTHORS: Anna Santin, Monia Teresa Russo, Pina Marotta, Francesco Manfellotto, Antonella Ruggiero, Mariella Ferrante [DESCRIPTION] Optimized protol to transform the pennate diatom Pseuodo-nitzschia multistriata. [STEPS] SECTION: First day: Cells...
["[First day: Cells plating] Plate 5 x 106 cells of P. multistriata culture in exponential phase (1.5-2 x 105 cells/ml) on 0.4% agarose in F/2 plates. Make a circular cell mat of diameter 4 cm.", "Sterilize rapture disks and macrocarriers by soaking for 15 min in 70% EtOH, then allow them to dry in sterile hood.", "[Be...
70,383
OT-2 PCR sample preparation protocol  
5
dx.doi.org/10.17504/protocols.io.n92ldpyznl5b/v1
https://www.protocols.io/view/ot-2-pcr-sample-preparation-protocol-cgyptxvn
Ana Mariya Anhel, Lorea Alejaldre, Manuel Gimenez, Ángel Goñi-Moreno
TITLE: OT-2 PCR sample preparation protocol   AUTHORS: Ana Mariya Anhel, Lorea Alejaldre, Manuel Gimenez, Ángel Goñi-Moreno [DESCRIPTION] This protocol is meant to perform samples preparation of PCR plates for several primer sets to the same samples, i.e, we will have for all the selected samples (we can set the initi...
["[Files Preparation] Preparing Customized Template\n\nPreparing the template (a .csv) with the specific variables for each experiment.\n\nHere we attach one excel with several sheets:\nTemplate to use in protocol\nExplanation of each variable\nSeveral examples", "[Running Protocol] Setting Labware", "[After-running] R...
63,843
Quant-IT DNA Quantification (Assay)
4
null
https://www.protocols.io/view/quant-it-dna-quantification-assay-cakbscsn
Allyson Hirsch, George Testo
TITLE: Quant-IT DNA Quantification (Assay) AUTHORS: Allyson Hirsch, George Testo [DESCRIPTION] Perform dsDNA quantification easily and quickly with Quant-iT dsDNA assay kits. Both the Quant-iT High-Sensitivity dsDNA Assay Kit and the Quant-iT Broad-Range dsDNA Assay Kit provide concentrated assay reagent, dilution bu...
["[Making Working Solution] Make working solution (number of samples + 24 standards + 10% = total volume).", "[Making Working Solution] 200uL Buffer (per sample) + 1uL Reagent (per sample)", "[Making Working Solution] Cover conical with a foil seal to prevent light exposure.", "[Making Working Solution] Vortex working ...
89,686
Protocol for Facially Guided Digital Diagnosis in Orthodontics
1
dx.doi.org/10.17504/protocols.io.8epv5x9q6g1b/v2
https://www.protocols.io/view/protocol-for-facially-guided-digital-diagnosis-in-c3twynpe
Rupert HG Kelley, Álvaro Ferrando Cascales, Raúl Ferrando Cascales
TITLE: Protocol for Facially Guided Digital Diagnosis in Orthodontics AUTHORS: Rupert HG Kelley, Álvaro Ferrando Cascales, Raúl Ferrando Cascales [DESCRIPTION] As the digital age of dentistry continues to flourish, it has never been more important to have protocols to guide dentists through the planning and performan...
["[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...
60,003
Shipping wastewater samples to FDA-CFSAN
1
dx.doi.org/10.17504/protocols.io.14egn797zv5d/v1
https://www.protocols.io/view/shipping-wastewater-samples-to-fda-cfsan-b6ubresn
Maria Balkey
TITLE: Shipping wastewater samples to FDA-CFSAN AUTHORS: Maria Balkey [DESCRIPTION] This SOP provides guidance for shipping wastewater samples to the US FDA Center for Food Safety and Applied Nutrition (CFSAN), covering specific steps for registering samples with CFSAN and for the preparation of shipments (documenta...
["[Submission of metadata] Fill out the BioSample custom wastewater template (extension of NCBI's Generic SARS-CoV-2: wastewater surveillance, v1.0 ) according to the NCBI submission protocol for SARS-Cov-2 protocol, once completed, send it to covidtrakr@fda.hhs.gov for registration. Successful registrations will re...
73,191
Staining of Gfap, Iba1,and NeuN on PFA-fixed mouse brain sections
1
dx.doi.org/10.17504/protocols.io.4r3l27q5pg1y/v1
https://www.protocols.io/view/staining-of-gfap-iba1-and-neun-on-pfa-fixed-mouse-cjqfumtn
Daniel Manrique-Castano
TITLE: Staining of Gfap, Iba1,and NeuN on PFA-fixed mouse brain sections AUTHORS: Daniel Manrique-Castano [DESCRIPTION] Staining protocol for Gfap, Iba1, and NeuN on PFA-fixed mouse brain sections. [GUIDELINES] Read the full protocol before starting the procedure. Note that this protocol uses 3 hours (room temperat...
["[Tissue preparation and blocking] Take out sections from -80 and place them in an incubator/plate for 20 min at 37 °C. This step is performed to ensure tissue attachment to the crystal slides.", "[Tissue preparation and blocking] 2. Draw a hydrophobic barrier on each slide using ImmEdge®Hydrophobic Barrier PAP Pen a...
60,419
Activation Induced Marker (AIM) Staining Protocol
4
dx.doi.org/10.17504/protocols.io.4r3l2o9w3v1y/v1
https://www.protocols.io/view/activation-induced-marker-aim-staining-protocol-b69brh2n
Gregory P. Williams, cecilia
TITLE: Activation Induced Marker (AIM) Staining Protocol AUTHORS: Gregory P. Williams, cecilia [DESCRIPTION] This protocol details about activation induced marker staining. [STEPS] SECTION: Peptide Stimulation Solution 1. Label U-bottom plate with donor, stimulation solution, name and date. SECTION: Peptide Stimulati...
["[Peptide Stimulation Solution] Label U-bottom plate with donor, stimulation solution, name and date.", "[Peptide Stimulation Solution] Prepare PHA and DMSO mix separately", "[Peptide Stimulation Solution] Prepare and arrange the remaining stimulation solution. Mix thoroughly by pipetting up and down before adding to ...
41,198
HTAPP_Dissociation of primary neuroblastoma core needle biopsy to a single-cell suspension for single-cell RNA-seq (using papain, density gradient, and optionally ACK)
1
dx.doi.org/10.17504/protocols.io.bkgnktve
https://www.protocols.io/view/htapp-dissociation-of-primary-neuroblastoma-core-n-bkgnktve
Sara Napolitano, Jingyi Wu, Michal Slyper, Sébastien Vigneau, Asaf Rotem, Bruce Johnson, Orit Rozenblatt-Rosen, Aviv Regev
TITLE: HTAPP_Dissociation of primary neuroblastoma core needle biopsy to a single-cell suspension for single-cell RNA-seq (using papain, density gradient, and optionally ACK) AUTHORS: Sara Napolitano, Jingyi Wu, Michal Slyper, Sébastien Vigneau, Asaf Rotem, Bruce Johnson, Orit Rozenblatt-Rosen, Aviv Regev [DESCRIPTIO...
["[Tissue Description]\nReport sample processing information.\nSample ID:Date:Time Received:Anatomical Site of the Biopsy:Number of Biopsy Cores:Core(s) Priority Number:Media Used for Transportation:Person Processing:", "[Tissue Description]\nTransfer sample to a Petri dish with cold PBS kept on ice to better visualize...
94,958
Assessing Growth Parameters in amphipod, Parhyale hawaiensis: Weighing and Measuring Length
4
dx.doi.org/10.17504/protocols.io.6qpvr3552vmk/v1
https://www.protocols.io/view/assessing-growth-parameters-in-amphipod-parhyale-h-c8ynzxve
Ibrahim Lawan
TITLE: Assessing Growth Parameters in amphipod, Parhyale hawaiensis: Weighing and Measuring Length AUTHORS: Ibrahim Lawan [DESCRIPTION] The developed protocol provides a standardised method for precisely determining the weight and length of Parhyale hawaiensis, a crucial model organism in ecotoxicology and ecophysiolo...
["[Introduction] Parhyale hawaiensis is a circumtropical marine amphipod crucial for epibenthic communities in tropical marine ecosystems (Poovachiranon et al., 1986). This amphipod has gained recognition for its sensitivity to environmental stressors and ecological significance, making it a promising model organism fo...
null
null
null
dx.doi.org/10.17504/protocols.io.im6cc9e
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>Southeast (SE) Asia is one of the most biodiverse regions in the world and it holds approximately 20% of all mammal species. Despite this, the majority of SE Asia’s genetic diversity is still poorly characterized. The growing interest in using environmental DNA (eDNA) to asse...
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86,796
The Anniversary Study – 20 years of Clavien-Dindo Classification &amp; 10 years of Comprehensive Complication Index®
1
dx.doi.org/10.17504/protocols.io.bp2l6xxzrlqe/v1
https://www.protocols.io/view/the-anniversary-study-20-years-of-clavien-dindo-cl-cyzkxx4w
Fariba Abbassi, Matthias Pfister, Anja Domenghino, Milo A Puhan, Pierre-A. Clavien
TITLE: The Anniversary Study – 20 years of Clavien-Dindo Classification &amp; 10 years of Comprehensive Complication Index® AUTHORS: Fariba Abbassi, Matthias Pfister, Anja Domenghino, Milo A Puhan, Pierre-A. Clavien [DESCRIPTION] Outcome reporting of surgical procedures was revolutionized in 2004 with the introducti...
["[Background] Outcome reporting of surgical procedures was revolutionized in 2004 with the introduction of the Clavien-Dindo (CD) classification.1 This 5-scale grading system offers an objective, simple, reliable, and reproducible way of recording complications, and has gained wide acceptance among the international s...
63,327
Trim Clinical Keto Review
3
dx.doi.org/10.17504/protocols.io.14egn7n5yv5d/v1
https://www.protocols.io/view/trim-clinical-keto-review-b937r8rn
JeffDlton
TITLE: Trim Clinical Keto Review AUTHORS: JeffDlton [DESCRIPTION] Trim Clinical Keto is a fat burner formula to get you a slim & trim figure Safe For You Buy Now On The Official Website [STEPS]
[]
71,490
Protocols from Kilfeather, Khoo et al., 2024
2
dx.doi.org/10.17504/protocols.io.36wgqj75kvk5/v1
https://www.protocols.io/view/protocols-from-kilfeather-khoo-et-al-2024-ch3at8ie
Peter Kilfeather, Maria Claudia Caiazza
TITLE: Protocols from Kilfeather, Khoo et al., 2024 AUTHORS: Peter Kilfeather, Maria Claudia Caiazza [DESCRIPTION] This protocol collection includes immunohisto/chemical staining methods, DAT-TRAP methods and the protocol for the Fura-2 assay in Kilfeather, Khoo et al., 2024. [STEPS]
[]
38,319
fatty acid analysis sponges
6
dx.doi.org/10.17504/protocols.io.bhnpj5dn
https://www.protocols.io/view/fatty-acid-analysis-sponges-bhnpj5dn
Anna de Kluijver
TITLE: fatty acid analysis sponges AUTHORS: Anna de Kluijver [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol has been used to analyse fatty acid composition of sponges (Porifera). The original protocol has been developed at NIOZ Yerseke, former NIOO-CEME (Netherlands) (Boschker and Mi...
["[Total lipid extraction]\nAdd Blight and Dyer mix, close with lid and shake/vortex\n28.5 mL", "[Total lipid extraction]\nWeight 30-50 mg dried sponge sample in extraction tubes (Kimax®centrifuge tube). This amount is enough to do either lipid-class or total lipid analysis, if you want to do both, weight a double a...
54,355
Bacterial isolation and genomic DNA extraction of Mycobacterium avium from pig lymph nodes
2
dx.doi.org/10.17504/protocols.io.bzbtp2nn
https://www.protocols.io/view/bacterial-isolation-and-genomic-dna-extraction-of-bzbtp2nn
Tetsuya Komatsu, Kenji Ohya, Justice Opare Odoi, Shota Suganuma, Kotaro Sawai, Takayuki Wada, Tomotada Iwamoto, Fumito Maruyama
TITLE: Bacterial isolation and genomic DNA extraction of Mycobacterium avium from pig lymph nodes AUTHORS: Tetsuya Komatsu, Kenji Ohya, Justice Opare Odoi, Shota Suganuma, Kotaro Sawai, Takayuki Wada, Tomotada Iwamoto, Fumito Maruyama [DESCRIPTION] Mycobacterium avium subsp. hominissuis (MAH) is one of the most impor...
[]
38,236
SEM Metal Sputter Coater User Guide
1
dx.doi.org/10.17504/protocols.io.yxmvmxjynl3p/v1
https://www.protocols.io/view/sem-metal-sputter-coater-user-guide-bhj4j4qw
Dakota Betz
TITLE: SEM Metal Sputter Coater User Guide AUTHORS: Dakota Betz [DESCRIPTION] How to use the SEM Metal Sputter Coater at SIO. [STEPS] 3. Turn on the Argon (gas cylinder in the corner of the room, the first one (check that it doesn't say nitrogen), turn the knob to the left just until you feel it give way and Ar start...
["Turn on the Argon (gas cylinder in the corner of the room, the first one (check that it doesn't say nitrogen), turn the knob to the left just until you feel it give way and Ar starts to flow).", "To put specimens in the sputter coater, WITH GLOVES, carefully open the canister lid and with your non-dominant hand pick ...
46,953
qPCR Primer Design
1
dx.doi.org/10.17504/protocols.io.br4hm8t6
https://www.protocols.io/view/qpcr-primer-design-br4hm8t6
Steven Burgess
TITLE: qPCR Primer Design AUTHORS: Steven Burgess [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Protocol for designing qPCR primers for analysis of transcript abundance using SYBR Green chemistry.</div><div class = "text-block"><span>There are many good guides about how to do this online such as <...
["[Identify target sequence]\nDecide on a gene of interest, download the transcript sequence in .gb format and upload to a new folder on Benchling for analysis.\nIf analyzing an endogenous (as compared to transgene), gene sequences can be obtained from public databases such as NCBI. For many plant genes, my preference ...
26,070
EASY AND INEXPENSIVE NUCLEIC ACID EXTRACTION PROTOCOL FOR RECALCITRANT SPECIES
null
dx.doi.org/10.17504/protocols.io.5pwg5pe
null
Lilian Matallana, Yusuf Kurt, William Kohlway, John Frampton, Ross Whetten
TITLE: EASY AND INEXPENSIVE NUCLEIC ACID EXTRACTION PROTOCOL FOR RECALCITRANT SPECIES AUTHORS: Lilian Matallana, Yusuf Kurt, William Kohlway, John Frampton, Ross Whetten [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Background</span><span>: Some of the major co...
["[Prepare lysate]\nThis section provides guidelines and instructions to isolate DNA and RNA from different types of tissues and plant species. A general protocol is described but users need to read carefully for specific instructions to improve the quality and yield of the isolated genomic material. We have tested bot...
19,744
U Mass - STZ-induced type 1 diabetes model
null
dx.doi.org/10.17504/protocols.io.xh8fj9w
null
Jason Kim
TITLE: U Mass - STZ-induced type 1 diabetes model 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">Streptozotocin can selectively destroy the pancreatic β-cell...
["Administer an intraperitoneal injection of streptozotocin (50 mg/kg body weight) daily for 5 days.", "Monitor glucose level for onset of hyperglycemia."]
41,765
Aegea Biotechnologies rapid PCR SARS-CoV-2 test (high sensitivity & specificity; able to detect different strain types)
4
dx.doi.org/10.17504/protocols.io.bk2dkya6
https://www.protocols.io/view/aegea-biotechnologies-rapid-pcr-sars-cov-2-test-hi-bk2dkya6
Lyle J. Arnold Ph.D., Stella M. Sung Ph.D.
TITLE: Aegea Biotechnologies rapid PCR SARS-CoV-2 test (high sensitivity & specificity; able to detect different strain types) AUTHORS: Lyle J. Arnold Ph.D., Stella M. Sung Ph.D. [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is for the Aegea Biotechnologies rapid PCR-based SARS-CoV-2...
["Prepare Reaction Mix10ul of the isolated total RNA will be used in a 20ul reaction with TaqPath™ 1-Step RT-qPCR Master Mix (5ul TaqPath Mastermix and 5ul of an oligo mix containing a. 0.5uM AEGEASwitch-Blocker (FAM),b. 0.5uM AEGEA COVID Forward Primer C, c. 2uM AEGEA COVID Reverse Primer, d. 0.8uM AEGEA L-Strain ...
36,739
Zooarchaeology by Mass Spectrometry (ZooMS) for bone material - Acid soluble protocol
1
dx.doi.org/10.17504/protocols.io.bf5bjq2n
https://www.protocols.io/view/zooarchaeology-by-mass-spectrometry-zooms-for-bone-bf5bjq2n
Samantha Brown, Sandra Hebestreit, Naihui Wang, Nicole Boivin, Katerina Douka, Kristine Richter
TITLE: Zooarchaeology by Mass Spectrometry (ZooMS) for bone material - Acid soluble protocol AUTHORS: Samantha Brown, Sandra Hebestreit, Naihui Wang, Nicole Boivin, Katerina Douka, Kristine Richter [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This collection details the different established prot...
["[Reagent preparation]\nGeneral Information:We recommend to use glassware for buffers and solutions and no long-term storage in plastic containers and/or tubes.TFA, HCl and ACN stock solutions should be handled under a fume hood only.Always label all chemicals with the compound name, concentraion and the date.Chemical...
81,531
An affordable solution for investigating zebra finch intracranial EEG signals
1
dx.doi.org/10.17504/protocols.io.q26g7y2w8gwz/v1
https://www.protocols.io/view/an-affordable-solution-for-investigating-zebra-fin-ctu3wnyn
Mohammad-Mahdi Abolghasemi, Shahrar Rezghi Shirsavar, Milad Yekani
TITLE: An affordable solution for investigating zebra finch intracranial EEG signals AUTHORS: Mohammad-Mahdi Abolghasemi, Shahrar Rezghi Shirsavar, Milad Yekani [DESCRIPTION] The Zebra finch is a well-studied model animal for neural mechanisms of speech, and Electrophysiology is the primary technique for understandi...
["[Hardware preparation] Electrode preparation:\nFor recording electrodes, we used 2.5 cm of stainless steel wire and uncoat two ends of it with a scalpel blade. \nmake a loop in one end about 1 or 2mm in diameter. use a surgical clamp to bend it. \nDo not make a bigger loop, it may not fit into the hole in the sk...
47,326
Immunofluorescence staining of PFA or fresh frozen mouse brain section
4
null
https://www.protocols.io/view/immunofluorescence-staining-of-pfa-or-fresh-frozen-bsf6nbre
Daniel Manrique-Castano
TITLE: Immunofluorescence staining of PFA or fresh frozen mouse brain section AUTHORS: Daniel Manrique-Castano [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is suitable for immunofluorescence staining of PFA fixed or Fresh Frozen mouse brain sections.</div></div> [STEPS] ?. [Rehydra...
["[Rehydration and permeabilization]\nIf fixed sections are kept in , rehydrate them before starting the staining procedure by incubating themin PBS for .\n-80 °C\nFor fresh frozen sections kept at , incubate with PFA during followed by 3 washes in PBS, each.", "[Rehydration and permeabilization]\nTo permeabi...
63,378
Where Can I Buy Ikaria Lean Belly Juice: Does It Work?? Must Know !!
3
dx.doi.org/10.17504/protocols.io.261genjp7g47/v1
https://www.protocols.io/view/where-can-i-buy-ikaria-lean-belly-juice-does-it-wo-b95sr86e
healthodiet
TITLE: Where Can I Buy Ikaria Lean Belly Juice: Does It Work?? Must Know !! AUTHORS: healthodiet [DESCRIPTION] Where Can I Buy Ikaria Lean Belly Juice: Does It Work?? Must Know !! [STEPS]
[]
61,564
Getting started with Micro-Meta App Tutorial
5
null
https://www.protocols.io/view/getting-started-with-micro-meta-app-tutorial-b8c4rsyw
Alessandro Rigano, Mathias Hammer, Joel Ryan, Claire M. Brown, David Grunwald, Caterina Strambio De Castillia
TITLE: Getting started with Micro-Meta App Tutorial AUTHORS: Alessandro Rigano, Mathias Hammer, Joel Ryan, Claire M. Brown, David Grunwald, Caterina Strambio De Castillia [DESCRIPTION] For quality, interpretation, reproducibility, and sharing value, microscopy images should be accompanied by detailed descripti...
["[Before the tutorial] Video introduction", "[Tutorial - 1 - Download and Install Micro-Meta App] Download Micro-Meta App\n\nFollow the instructions in this step and in Video 3 of the tutorial series to download and install the stand-alone version of the Micro-Meta App.", "[Tutorial - 1 - Download and Install Micro-Me...
61,921
Custom Peptide Design
6
dx.doi.org/10.17504/protocols.io.ewov1n35ygr2/v1
https://www.protocols.io/view/custom-peptide-design-b8p9rvr6
Cathy Miller
TITLE: Custom Peptide Design AUTHORS: Cathy Miller [DESCRIPTION] Interested in the use of biologically active proteins and peptides as potential therapeutic agents has grown dramatically in recent years. Although many meaningful studies can be performed, it is important to consider several factors such as the length ...
[]
44,232
Stable transfection of unicellular relative of animals, Corallochytrium limacisporum, using Lonza Nucleofector
1
null
https://www.protocols.io/view/stable-transfection-of-unicellular-relative-of-ani-bpfgmjjw
Aleksandra Kozyczkowska, Sebastian Najle, Eduard Ocaña-Pallarès, Cristina Aresté, Iñaki Ruiz-Trillo, Elena Casacuberta
TITLE: Stable transfection of unicellular relative of animals, Corallochytrium limacisporum, using Lonza Nucleofector AUTHORS: Aleksandra Kozyczkowska, Sebastian Najle, Eduard Ocaña-Pallarès, Cristina Aresté, Iñaki Ruiz-Trillo, Elena Casacuberta [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span...
["[Cell Culture preparation ]\nInoculate 100 μl of C. limacisporum culture in 5ml of Marine Broth in 25cm2 flask. Keep at 23°C air incubator for two days.", "[Cell count]\nScrape gently and count cells to reach 1.5E6 cells / condition using hemocytometerIf there are more conditions, it is recommended to pull the cells ...
37,930
Perceptions, knowledge and attitudes of the adult populations towards COVID-19 : A scoping review protocol
1
dx.doi.org/10.17504/protocols.io.bhaij2ce
https://www.protocols.io/view/perceptions-knowledge-and-attitudes-of-the-adult-p-bhaij2ce
Nathalie Clavel, Mathieu Seppey, Lara Gautier, Mélanie Lavoie-Tremblay
TITLE: Perceptions, knowledge and attitudes of the adult populations towards COVID-19 : A scoping review protocol AUTHORS: Nathalie Clavel, Mathieu Seppey, Lara Gautier, Mélanie Lavoie-Tremblay [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Perceptions, knowledge and attitudes of the adult populati...
["[Title and author identification]\nPerceptions, knowledge and attitudes of the adult populations towards COVID-19 : A scoping review protocol Nathalie Clavel1, Mathieu Seppey2, Lara Gautier2,3, Mélanie Lavoie-Tremblay11 Ingram School of Nursing, McGill University2 School of Public Health, University of Montreal 3 Dep...
54,816
Diagnostic accuracy of home sleep apnea testing using peripheral arterial tonometry for sleep apnea syndrome: protocol for a systematic review and meta-analysis
1
dx.doi.org/10.17504/protocols.io.bzr8p59w
https://www.protocols.io/view/diagnostic-accuracy-of-home-sleep-apnea-testing-us-bzr8p59w
Masahiro Ichikawa, Tomoaki Akiyama, Yasushi Tsujimoto, Keisuke Anan, Tadashi Yamakawa, Yasuo Terauchi
TITLE: Diagnostic accuracy of home sleep apnea testing using peripheral arterial tonometry for sleep apnea syndrome: protocol for a systematic review and meta-analysis AUTHORS: Masahiro Ichikawa, Tomoaki Akiyama, Yasushi Tsujimoto, Keisuke Anan, Tadashi Yamakawa, Yasuo Terauchi [DESCRIPTION] INTRODUCTION Ratio...
[]
75,908
Protocol: Neutrophil/lymphocyte ratio and overall survival in patients with breast cancer: a cohort study in a Latin-American hospital
1
dx.doi.org/10.17504/protocols.io.kxygx95xdg8j/v1
https://www.protocols.io/view/protocol-neutrophil-lymphocyte-ratio-and-overall-s-cndcva2w
Martha Sofia Cervera-Ocaña, Jhony A. De La Cruz-Vargas, Dante Manuel Quiñones-Laveriano, Nataly Briyit Huamán Córdova
TITLE: Protocol: Neutrophil/lymphocyte ratio and overall survival in patients with breast cancer: a cohort study in a Latin-American hospital AUTHORS: Martha Sofia Cervera-Ocaña, Jhony A. De La Cruz-Vargas, Dante Manuel Quiñones-Laveriano, Nataly Briyit Huamán Córdova [DESCRIPTION] PURPOSE OF STUDY The primary objecti...
["[Neutrophil/lymphocyte ratio and overall survival in patients with breast cancer: a cohort study in a Latin-American hospital]"]
106,575
MRI_preprocessing_protocol
0
dx.doi.org/10.17504/protocols.io.14egn61byl5d/v1
https://www.protocols.io/view/mri-preprocessing-protocol-dkbp4smn
Miguel López-Aguirre, José A. Pineda-Pardo, Jose Obeso
TITLE: MRI_preprocessing_protocol AUTHORS: Miguel López-Aguirre, José A. Pineda-Pardo, Jose Obeso [DESCRIPTION] Preprocessing protocol for structural, DWI and T2* MRI acquisitions [STEPS] SECTION: Structural T1w (MP-RAGE acquisition) 1. Data transformation from DICOM to NIFTI standard (software: dcm2niix) SECTION: S...
["[Structural T1w (MP-RAGE acquisition)] Data transformation from DICOM to NIFTI standard (software: dcm2niix)", "[Structural T1w (MP-RAGE acquisition)] Field of view optimization (command: fslreorient2std; software: FSL)", "[Structural T1w (MP-RAGE acquisition)] Bias field correction (command: N4BiasFieldCorrection; s...
41,075
TMA-TNP Section Map and Slide Processing - Phase 1
1
dx.doi.org/10.17504/protocols.io.bkctkswn
https://www.protocols.io/view/tma-tnp-section-map-and-slide-processing-phase-1-bkctkswn
Heidi Feiler, Koei Chin
TITLE: TMA-TNP Section Map and Slide Processing - Phase 1 AUTHORS: Heidi Feiler, Koei Chin [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Human Tumor Atlas Tissue MicroArrary TNP (TMA-TNP)</div><br/><div class = "text-block">The Tissue MicroArray (TMA) TNP will extend the SARDANA-TNP characteriz...
["[Preparation]\nVerify the identity of the FFPE block to be cut against written request for sectioning. The FFPE block (TMA1) will be utilized for TMA-TNP Phase 1.", "[Preparation]\nEach slide was labeled with a unique OHSU Slide ID corresponding to the FFPE section map (below). ABCDE1&nbsp;Slide ID&nbsp;&nbsp;&nbs...
63,348
Production: 10x TBE Buffer Powder Sachets
4
null
https://www.protocols.io/view/production-10x-tbe-buffer-powder-sachets-b94ur8ww
Jenny Molloy, Nadine Mowoh, Stephane Fadanka
TITLE: Production: 10x TBE Buffer Powder Sachets AUTHORS: Jenny Molloy, Nadine Mowoh, Stephane Fadanka [DESCRIPTION] Desired concentrations of Components in the 10x buffer Boric acid-0.89M Tris-0.89M Na2EDTA-0.02M Concentration of 1x TBE buffer Tris-Borate-0.089M Na2EDTA-0.002M [BEFORE_START] Ensure all powder...
["Weigh about 35 g of Boric acid into a weighing boat and place the weighing boat in a vacuum tupperware half-filled with silica gel beads.", "Close the tupperware and use the vacuum pump to create a vacuum until you are not able to hand pump any more air out or the pump shuts off.", "Allow it to stay overnight at room...
95,960
Western blot for tissue extract
0
dx.doi.org/10.17504/protocols.io.yxmvm38q9l3p/v1
https://www.protocols.io/view/western-blot-for-tissue-extract-c9xyz7pw
María Sanchiz Calvo, eduard.bentea, Veerle Baekelandt
TITLE: Western blot for tissue extract AUTHORS: María Sanchiz Calvo, eduard.bentea, Veerle Baekelandt [DESCRIPTION] Protocol for the detection of proteins by Western blot from tissue extract [BEFORE_START] Perform protein extraction from snap-frozen brain tissue: - weigh tissue - add RIPA buffer (see Materials) : 1...
["[Day 1] Prepare sample for western blot\n15 µg of protein in 12 µL \nAdd 4 µL \nBoil at 98 °C for 10 min", "[Day 1] Load samples, together with 7 µL of mass marker (PageRuler‱ Plus Prestained Protein Ladder) on a on 4–15% Criterion‱ Tris-HCl Protein Gel.", "[Day 1] Run the gel for 10 min at 80V", "[Day 1] Run the g...
86,602
U54 SCENT Intracellular Staining (ICS) Senescence Flow Cytometry Panel
1
dx.doi.org/10.17504/protocols.io.rm7vzxxk5gx1/v1
https://www.protocols.io/view/u54-scent-intracellular-staining-ics-senescence-fl-cytixwke
Zachary R Healy, David Murdoch, Alicia Cooper-Volkheimer
TITLE: U54 SCENT Intracellular Staining (ICS) Senescence Flow Cytometry Panel AUTHORS: Zachary R Healy, David Murdoch, Alicia Cooper-Volkheimer [DESCRIPTION] This protocol describes the Intracellular Staining (ICS) Senescence Flow Cytometry Panel [STEPS] SECTION: FBS Aliquot Prep 1. FBS (hiFBS): Gemini Bio Products ...
["[FBS Aliquot Prep] FBS (hiFBS): Gemini Bio Products Cat #100-106 1L\nPrepare FBS Aliquots:", "[FBS Aliquot Prep] Thaw heat-inactivated FBS (hiFBS):\nThaw a 500mL Bottle of FBS at 4°C. This may require more than overnight so the 500mL Bottle may be removed 2 or 3 days prior to use. Do not leave the FBS at room tempera...
67,728
Visualization of a low concentration and molecular weight DNA (DNA gel stain)
4
dx.doi.org/10.17504/protocols.io.5jyl89x46v2w/v2
https://www.protocols.io/view/visualization-of-a-low-concentration-and-molecular-cedqta5w
Nadine Mowoh, Stephane Fadanka, Shalo Minette
TITLE: Visualization of a low concentration and molecular weight DNA (DNA gel stain) AUTHORS: Nadine Mowoh, Stephane Fadanka, Shalo Minette [DESCRIPTION] After PCR amplification of DNA, agarose gel electrophoresis is run to separate the DNA based on their size. The agarose gel consists of microscopic pores ...
["[Visualization of a low concentration and molecular weight DNA]", "[Visualization of a low concentration and molecular weight DNA] To confirm visualization of a low molecular weight DNA\n\n\nFollow the steps in preparing a 2% agarose gel as described in this protocol, \n \n\nLoad the gel (made from the test gel stain...
30,052
PROTOCOL FOR LARVICIDE BIOASSAYS
null
dx.doi.org/10.17504/protocols.io.9kch4sw
null
Dr. Wanderli Pedro Tadei, Rochelly da Silva Mesquita
TITLE: PROTOCOL FOR LARVICIDE BIOASSAYS AUTHORS: Dr. Wanderli Pedro Tadei, Rochelly da Silva Mesquita [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol is designed for the evaluation of the larvicidal activity of perspective insecticides on Anopheles larvae. The tests are performed accor...
["Preparation of the stock solutionsDissolve accurately weighed quantity of the test substance in an appropriate volume of dimethylsulfoxide (DMSO) and/or distilled water (depends on the sample solubility) to obtain the concentration of .Final volume suggestion: to , depending on the amount of the material available ...
11,024
Thigmotaxis_Detection_2018
null
dx.doi.org/10.17504/protocols.io.nzqdf5w
null
Akinori Higaki, Masaki Mogi, Jitsuo Higaki, Masatsugu Horiuchi
TITLE: Thigmotaxis_Detection_2018 AUTHORS: Akinori Higaki, Masaki Mogi, Jitsuo Higaki, Masatsugu Horiuchi [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">We share the minimal dataset for our study "Recognition for Early Stage Thigmotaxis in Morris Water Maze Test with Convolutional Neural Network" h...
[]
35,809
HuBMAP UF TMC - 10x Genomics scRNAseq Modality Overview
null
dx.doi.org/10.17504/protocols.io.be79jhr6
https://www.protocols.io/view/hubmap-uf-tmc-10x-genomics-scrnaseq-modality-overv-be79jhr6
Marda Jorgensen, Maigan Brusko
TITLE: HuBMAP UF TMC - 10x Genomics scRNAseq Modality Overview AUTHORS: Marda Jorgensen, Maigan Brusko [STEPS] ?. Specimen/Organ Prep:Spleen: dx.doi.org/10.17504/protocols.io.bc3kiykwLymph Node: dx.doi.org/10.17504/protocols.io.bbgnijveThymus: dx.doi.org/10.17504/protocols.io.bbgmiju6 ?. Single Cell Isolation/Storage...
["Specimen/Organ Prep:Spleen: dx.doi.org/10.17504/protocols.io.bc3kiykwLymph Node: dx.doi.org/10.17504/protocols.io.bbgnijveThymus: dx.doi.org/10.17504/protocols.io.bbgmiju6", "Single Cell Isolation/Storage: dx.doi.org/10.17504/protocols.io.bd9vi966", "10x Library Construction and Sequencing: dx.doi.org/10.17504/protoc...
68,197
CompetentCell
4
dx.doi.org/10.17504/protocols.io.j8nlkkzd6l5r/v1
https://www.protocols.io/view/competentcell-ceudtes6
Ana Belem García González, Georgina Diego, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz
TITLE: CompetentCell AUTHORS: Ana Belem García González, Georgina Diego, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz [DESCRIPTION] A protocol for making chemically competent cells is shown. [STEPS] SECTION: Competent cells protocol with CaCl2 1. Inoculate 100 µL of E. coli in10 mL of antibiotic-free...
["[Competent cells protocol with CaCl2] Inoculate 100 µL of E. coli in10 mL of antibiotic-free starter LB culture medium. Incubate during at 37 °C and", "[Competent cells protocol with CaCl2] Heat the LB media in the water bath at37 °C", "[Competent cells protocol with CaCl2] Inoculate 40 mL of LB media with 1 mL of c...
58,991
623.1.HTC_Precision_Cut_Lung_Slices
1
dx.doi.org/10.17504/protocols.io.4r3l2o26xv1y/v1
https://www.protocols.io/view/623-1-htc-precision-cut-lung-slices-b5upq6vn
Cory Poole, Lisa Rogers, Gloria S Pryhuber
TITLE: 623.1.HTC_Precision_Cut_Lung_Slices AUTHORS: Cory Poole, Lisa Rogers, Gloria S Pryhuber [DESCRIPTION] Purpose and Scope of the Procedure or Laboratory Assay Provide high quality, viable, 500 µm sections of human lung for culture and other in vitro assays Current preparation has limited ability to produce qualit...
["[Inflation of the Lung] Before dissections begin, place new sterile HBSS in a clean water bath set between 37°C and 40°C. This way the HBSS will be up to temperature by the time inflations begin. Pour more HBSS into another clean container, that is places within another larger container filled with wet ice, that will...
null
null
null
dx.doi.org/10.17504/protocols.io.cjvun5
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?.
[]
99,144
Wake Forest University Health Sciences Manual of Procedures Biospecimen Collection and Processing V3
1
dx.doi.org/10.17504/protocols.io.kqdg32p6ev25/v1
https://www.protocols.io/view/wake-forest-university-health-sciences-manual-of-p-dc3g2yjw
Kim Kennedy, Heather Gregory
TITLE: Wake Forest University Health Sciences Manual of Procedures Biospecimen Collection and Processing V3 AUTHORS: Kim Kennedy, Heather Gregory [DESCRIPTION] This includes the methods associated with obtaining muscle, blood, and urine along with demographic, clinical, health, and functional data from younger and old...
[]
97,923
cosy_metab.nan
5
dx.doi.org/10.17504/protocols.io.eq2lyw3zpvx9/v1
https://www.protocols.io/view/cosy-metab-nan-dbvb2n2n
NAN KB, John Glushka, Mario Uchimiya, Saraa Al Jawad, Christopher Esselman, Leandro I Ponce, Laura Morris, Arthur Edison
TITLE: cosy_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 "cosygpprqf" for metabolomics samples. [BEFORE_START] This protocol assumes: Your sample is load...
["[Create a new dataset]", "[Create noesypr1d experiment file] When Lock, Tune, Shim are complete proceed to the specific protocol for your desired pulseprogram(s).", "[Create a new dataset] On the menu bar on TopSpin, click on\nStart → Create Dataset", "[Create a new dataset] Enter\nNAME: Name of a set of datasets (e....
91,406
Preparation of cells for live-cell imaging of a FOXO-based AKT kinase translocation reporter by widefield microscopy
4
dx.doi.org/10.17504/protocols.io.261gedjkjv47/v1
https://www.protocols.io/view/preparation-of-cells-for-live-cell-imaging-of-a-fo-c5hny35e
Ralitsa R Madsen
TITLE: Preparation of cells for live-cell imaging of a FOXO-based AKT kinase translocation reporter by widefield microscopy AUTHORS: Ralitsa R Madsen [DESCRIPTION] This protocol provides detailed instructions for imaging of cells with stable expression of a kinase translocation reporter, alongside a nuclear marker for...
["[Dish assembly and coating] Take the required number of Ibidi dishes that will be needed for the experiment, along with the equivalent number of inserts. (NB: you can reuse inserts once provided that they have been sterilised in 70% ethanol and dried since the last use; we do this by passing them through PBS once, th...
91,885
Human iPSCs culture and cardiomyocyte subtype differentiation in fully chemically defined conditions 
4
null
https://www.protocols.io/view/human-ipscs-culture-and-cardiomyocyte-subtype-diff-c5ymy7u6
Joanna Delimata-Raczek, Natalia Koralewska, Marek Figlerowicz, Ireneusz Stolarek
TITLE: Human iPSCs culture and cardiomyocyte subtype differentiation in fully chemically defined conditions  AUTHORS: Joanna Delimata-Raczek, Natalia Koralewska, Marek Figlerowicz, Ireneusz Stolarek [DESCRIPTION] Cardiomyocyte in vitro differentiation represents a pivotal avenue in stem cell research, offering unprece...
["[iPSC culture and maintenance] iPSC culture", "[Subtype differentiation of hiPSCs into cardiomyocytes] Standard iPSC culture before differentiation.", "[iPSC culture and maintenance] iPSC passage", "[iPSC culture and maintenance] iPSC cryopresevation", "[iPSC culture and maintenance] The optimal time for harvest or p...
50,463
Making normal NGM
1
dx.doi.org/10.17504/protocols.io.bvh7n39n
https://www.protocols.io/view/making-normal-ngm-bvh7n39n
Bonnie Evans
TITLE: Making normal NGM AUTHORS: Bonnie Evans [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">C. elegans is maintained in the laboratory on Nematode Growth Medium (NGM) agar which has been aseptically poured into petri plates using a peristaltic pump. NGM (rather than no peptone NGM) may also be us...
["[Pouring multi-well plates]\nFor pouring multi-well imaging plates:\n{\"blocks\":[{\"key\":\"fmd1o\",\"text\":\"Protocol for pouring agar into 96 well plates using Integra VIAFILL dispenser. Agar should be prepared in advance, and kept in 60oC waterbath until ready to dispense, and whilst dispensing. The X, Y, Z posi...
107,129
Single_Stranded_Library_Workflow_Gut_Virome
0
dx.doi.org/10.17504/protocols.io.8epv5r5w5g1b/v1
https://www.protocols.io/view/single-stranded-library-workflow-gut-virome-dkuz4wx6
Xichuan Zhai
TITLE: Single_Stranded_Library_Workflow_Gut_Virome AUTHORS: Xichuan Zhai [DESCRIPTION] Protocol of single-stranded library (SSLR) preparation for virome study [BEFORE_START] Fill bucket with crushed ice. [GUIDELINES] This protocol is designed for virome library preparation with our lab-customed single-stranded libra...
["[Step1: Virome isolation and extraction] Checklist before starting", "[Step2: Heat treatment] Checklist before starting", "[Step3: Reverse transcription (RT)] Checklist before starting", "[Step4: Fragmentation] Checklist before starting", "[Step5: Ligation] Checklist before starting", "[Step6: Index PCR] Checklist be...
41,923
Acute toxicity of injected drugs and substances in fish
4
dx.doi.org/10.17504/protocols.io.bk7bkzin
https://www.protocols.io/view/acute-toxicity-of-injected-drugs-and-substances-in-bk7bkzin
Bruna Patrícia D.c., Sabrina Alana Gomes Pinto, Layana Aquino Moura, Diógenes Silva, Kelly Christina Ferreira Castro, Caio Maximino
TITLE: Acute toxicity of injected drugs and substances in fish AUTHORS: Bruna Patrícia D.c., Sabrina Alana Gomes Pinto, Layana Aquino Moura, Diógenes Silva, Kelly Christina Ferreira Castro, Caio Maximino [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">This protocol, modified from OECD 203 (Acute to...
["[Anesthesia and injection procedures]\nAnesthesize fish in ice-cold water (12-14 ºC). Remove animal from water as soon as movements stop.\non ice", "[Anesthesia and injection procedures]\nTransfer animal to surgical bed (water-soaked sponge with a cut to fixate the fish), kept in a vessel with ice-cold water.\non ice...
23,874
BGISEQ-500 10X library construction
null
dx.doi.org/10.17504/protocols.io.3jagkie
null
ziqiang chen
TITLE: BGISEQ-500 10X library construction AUTHORS: ziqiang chen [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">BGISEQ-500 is a desktop sequencer developed by BGI. This protocol adjusts the process in order to apply the 10X contruction to the BGISEQ-500 rather than illumina.</div></div> [STEPS] ?....
["Input HMW gDNA Quantification", "Quantitate 3 μl of extracted gDNA solution (with a minimum of 2 replicates).", "If the gDNA stock is >20 ng/μl, prepare an intermediate dilution of the extracted gDNA solution at", "Quantitate 3 μl of the", "According to the table below, dilute gDNA with bufferEB concentration less th...
35,379
Opentrons COVID-19 testing (Zymo, station B, 48 samples)
null
dx.doi.org/10.17504/protocols.io.bestjeen
null
Max Marrone
TITLE: Opentrons COVID-19 testing (Zymo, station B, 48 samples) AUTHORS: Max Marrone [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Station B: Initial OT-2 setup] Clean the OT-2. ?. [Station B: Initial OT-2 setup] Place the following labware on the OT-2's deck:Slot 1: An Opentrons Temperature Module wit...
["[Station B: Initial OT-2 setup]\nClean the OT-2.", "[Station B: Initial OT-2 setup]\nPlace the following labware on the OT-2's deck:Slot 1: An Opentrons Temperature Module with an Opentrons 96 well aluminum block and an empty, sterile NEST 100 µL PCR plate.Slots 3, 6, 7, 9, and 10: A full, sterile rack of Opentrons 2...
57,737
Ultra-high field, multi-echo MRI acquisition for resting-state functional connectivity
1
dx.doi.org/10.17504/protocols.io.dm6gpbbj1lzp/v1
https://www.protocols.io/view/ultra-high-field-multi-echo-mri-acquisition-for-re-b4mhqu36
Elizabeth Rizor, Dr. Scott Grafton
TITLE: Ultra-high field, multi-echo MRI acquisition for resting-state functional connectivity AUTHORS: Elizabeth Rizor, Dr. Scott Grafton [DESCRIPTION] This is an MRI acquisition sequence used to examine resting-state functional connectivity of open and closed loop motor circuits in healthy controls and those with Par...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.jqjcmun
null
null
TITLE: No Title AUTHORS: [STEPS] ?. ?. ?. ?. ?. ?.
[]
null
null
null
dx.doi.org/10.17504/protocols.io.ekwbcxe
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] Stellaris® RNA FISH Cells in Suspension Protocol is specifically optimized to allow to visualization of single RNA transcripts in suspended, fixed cells. [BEFORE_START] <p align="LEFT">Reagents and Equipment</p> <p align="LEFT">Reagents and Consumables:</p> <p align="LEFT">a) T...
[]
18,493
Water Sampling and Plating Generations (Antimicrobial Group)
1
dx.doi.org/10.17504/protocols.io.ewov1yd87vr2/v1
https://www.protocols.click/view/water-sampling-and-plating-generations-antimicrobi-wa5fag6
Melissa B Duhaime, Aubrey Reed, Nicole Sunshine, Renae Lyons
TITLE: Water Sampling and Plating Generations (Antimicrobial Group) AUTHORS: Melissa B Duhaime, Aubrey Reed, Nicole Sunshine, Renae Lyons [DESCRIPTION] Protocol developed by the Antimicrobial Group class project. The goal was to isolate microbes from freshwater lakes that produce antimicrobial compounds in order to st...
["Collect whole water at six sites using a Van Dorn water collector for deep (~122.5-123.7 m) samples and 2 L containers for surface samples (~15 cm below the surface).", "Set site names and classifications\n \n3. Prefilter whole water using 20μm mesh. \n4. Set up peristaltic pump with 3 µm filter\n5. Filter approxima...
91,600
The relationship between chronic skin conditions and depression and how successful dermatologic treatment could improve patients’ mental health: An evidence-based systematic review protocol
1
null
https://www.protocols.io/view/the-relationship-between-chronic-skin-conditions-a-c5pqy5mw
Aime Lee, Nami Watanabe, Emma Gregory, Cobi Desilva, Jenina Joyce Pali, Lindsey Leonhard, Samia Valeria Ozorio Dutra
TITLE: The relationship between chronic skin conditions and depression and how successful dermatologic treatment could improve patients’ mental health: An evidence-based systematic review protocol AUTHORS: Aime Lee, Nami Watanabe, Emma Gregory, Cobi Desilva, Jenina Joyce Pali, Lindsey Leonhard, Samia Valeria Ozorio Dut...
[]
94,688
Rotarod test in rats
1
dx.doi.org/10.17504/protocols.io.5qpvo3zo9v4o/v1
https://www.protocols.io/view/rotarod-test-in-rats-c8p8zvrw
eduard.bentea, María Sanchiz Calvo, Veerle Baekelandt
TITLE: Rotarod test in rats AUTHORS: eduard.bentea, María Sanchiz Calvo, Veerle Baekelandt [DESCRIPTION] Protocol for performing the rotarod test in rats. This test evaluated motor coordination and balance, and can be used in rat models with motor deficits, such as models of Parkison's disease. Rats with impaired moto...
["[Training] Bring cages to the behavioral room for at least 60 min prior to the test for habituation", "[Training] Habituate rats on the rotarod for 5 min at 5 rpm, place back in case they fall", "[Training] 5 min rest", "[Training] Test rats for 1 min at 5 rpm", "[Training] 5 min rest", "[Training] Test rats for 1 mi...
94,117
Immunohistochemistry free-floating rat brain cryosections  
4
dx.doi.org/10.17504/protocols.io.3byl4q4zrvo5/v1
https://www.protocols.io/view/immunohistochemistry-free-floating-rat-brain-cryos-c76dzra6
mariangela.massarocenere
TITLE: Immunohistochemistry free-floating rat brain cryosections   AUTHORS: mariangela.massarocenere [DESCRIPTION] Protocol for immunohistochemistry on rat brain cryosections [STEPS] SECTION: 1. Sections selection 1. Collect the cryosections needed for a caudo-rostral representation of each brain region (every fift...
["[1. Sections selection] Collect the cryosections needed for a caudo-rostral representation of each brain region (every fifth or sith sections depending on the section's thickness, brain region, and animal species) into a 24-well-plate (3-4 sections per well)", "[2. Inactivation of endogenous peroxidase] Incubate sec...
102,750
In vitro co-culture system using a fiber-supported liquid approach
0
dx.doi.org/10.17504/protocols.io.rm7vzj56xlx1/v1
https://www.protocols.io/view/in-vitro-co-culture-system-using-a-fiber-supported-dgj63ure
Alejandro Calle, Jeffrey Adelberg, Guido Schnabel, Jacqueline Naylor-Adelberg, Jhulia Gelain, Yeter Karakoc, Jared Weaver, Christopher Saski, Ksenija Gasic
TITLE: In vitro co-culture system using a fiber-supported liquid approach AUTHORS: Alejandro Calle, Jeffrey Adelberg, Guido Schnabel, Jacqueline Naylor-Adelberg, Jhulia Gelain, Yeter Karakoc, Jared Weaver, Christopher Saski, Ksenija Gasic [DESCRIPTION] In vitro co-culture techniques that allow the growth of plants and...
["[Establishment of Plant Cultures] Establishment of plant cultures from dormant shoots", "[Establishment of Plant Cultures] Collect dormant shoots, cut them (3 cm in length),and cleanse them by submerging in 70% ethanol for 1 minute, followed by rinsing with sterile deionized water. Then, immerse the shoots in a 10% b...
24,178
Podocyte Count and Density Analysis
null
dx.doi.org/10.17504/protocols.io.3usgnwe
null
Frank Brosius
TITLE: Podocyte Count and Density Analysis AUTHORS: Frank Brosius [DESCRIPTION] <div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This protocol describes the procedures for quantifying the podocyte count and density in the glomerulus....
["[Procedure:]\n1)  Stain the Perfusion-fixed paraffin embedded sections (3.9 and 10.2 microns thick) with WT-1 antibody and immunoperoxidase and then take photograph at 40x by using Spot Advanced Software Camera.\n2)  Photograph 50 consecutive glomerular cross-sections moving systematically from outer cortex to inner ...
51,934
TEA-seq
4
dx.doi.org/10.17504/protocols.io.bwx6pfre
https://www.protocols.io/view/tea-seq-bwx6pfre
Elliott Swanson, Lucas Graybuck, Peter Skene
TITLE: TEA-seq AUTHORS: Elliott Swanson, Lucas Graybuck, Peter Skene [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">TEA-seq is a method for Transcriptomic, Epitope, and Accessibility measurement from thousands of single cells on the 10x Genomics Multiome platform. It generates scRNA-seq, scATAC-seq...
["[Buffer preparation]\nStain Buffer Dulbecco's phosphate-buffered saline (DPBS) supplemented with 2% w/v bovine serum albumin.Wash Buffer Final composition of 20 mM Tris HCl (Tris(hydroxymethyl)aminomethane hydrochloride) pH 7.4, 150 mM NaCl, 3 mM MgCl2.Perm Buffer Wash Buffer (above) with the addition of digitonin t...
90,854
DNA barcoding SOPs for the Darwin Tree of Life Project
2
dx.doi.org/10.17504/protocols.io.261ged91jv47/v1
https://www.protocols.io/view/dna-barcoding-sops-for-the-darwin-tree-of-life-pro-c4yeyxte
Jordan Beasley, Rebekka Uhl, Laura L Forrest, David Bell, Michelle Hart, Rebecca Yahr, Estelle Kilias
TITLE: DNA barcoding SOPs for the Darwin Tree of Life Project AUTHORS: Jordan Beasley, Rebekka Uhl, Laura L Forrest, David Bell, Michelle Hart, Rebecca Yahr, Estelle Kilias [DESCRIPTION] This collection of protocols describe the standard operating procedures for DNA barcoding in the Darwin Tree of Life project. The SO...
[]
null
null
null
dx.doi.org/10.17504/protocols.io.rbsd2ne
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p>This protocol outlines the steps taken to support sexual reproduction in Aiptasia lab cultures. It also provides steps for handling embryos and larvae, and basic husbandry to keep them alive.</p> <p>Spawning protocol based on the methods from Grawunder et al. 2015.</p> [STEP...
[]
101,615
Single-step assembly of double guide plasmid (pCas9-Duo) for gene-editing in Plasmodium 
0
dx.doi.org/10.17504/protocols.io.n2bvjn9d5gk5/v1
https://www.protocols.io/view/single-step-assembly-of-double-guide-plasmid-pcas9-dfgp3jvn
Abhinay Ramaprasad
TITLE: Single-step assembly of double guide plasmid (pCas9-Duo) for gene-editing in Plasmodium  AUTHORS: Abhinay Ramaprasad [DESCRIPTION] This protocol describes a one-pot GoldenGate assembly of a new dual-guide targeting plasmid (pCas9-Duo) expressing two distinct guide RNAs in order to enhance the chances of a succe...
["[gRNA oligo design] Add the overhangs \"ATTG\" and \"AAAC\" to forward and reverse oligos of gRNA1, and \"TTGG\" and \"TAAA\" to gRNA2 respectively.", "[Anneal gRNA oligos] Set up annealing reactions. \n \n1.0 µL gRNA.F 100 micromolar (µM) \n1.0 µL gRNA.R 100 micromolar (µM) \n1.0 µL \n0.5 µL \n6.5 µL", "[An...
86,286
Live Imaging of Primary Mouse Neuron Cultures
4
dx.doi.org/10.17504/protocols.io.e6nvwdjydlmk/v1
https://www.protocols.io/view/live-imaging-of-primary-mouse-neuron-cultures-cyhnxt5e
Robert Edwards, Shweta Jain
TITLE: Live Imaging of Primary Mouse Neuron Cultures AUTHORS: Robert Edwards, Shweta Jain [DESCRIPTION] This protocol describes live imaging of primary neuron cultures. Included are methods for preparing hippocampal or dopamine neuronal cultures from neonatal mouse brain tissue. The imaging described involves labeling...
["[Preparation of Hippocampal Cultures] Collect mouse pups at postnatal day 0", "[Preparation of Hippocampal Cultures] Decapitate and remove brain, dissect out hippocampi from each hemisphere into Hank's balanced salt solution (HBSS) containing 10 mM HEPES and 20 mM glucose", "[Preparation of Hippocampal Cultures] Dige...
64,888
PLATERO - Green Fluorescence Calibration in Plate Readers
1
null
https://www.protocols.io/view/platero-green-fluorescence-calibration-in-plate-re-cbkyskxw
Alejandro Vignoni, Yadira Boada
TITLE: PLATERO - Green Fluorescence Calibration in Plate Readers AUTHORS: Alejandro Vignoni, Yadira Boada [DESCRIPTION] One of the most common sources of information in Synthetic Biology is the data coming from plate reader fluorescence measurements. These experiments provide a measure of the light emitted by certa...
["[Stock Reference Fluorescein solution] Start from at least 1 mL of 10 micromolar (µM) solution in .", "[Prepare the serial dilutions of Fluorescein] Accurate pipetting is essential. Serial dilutions will be performed in 5 tubes. There will be a sixth tube that must contain PBS buffer only. Initially, you will s...
null
null
null
dx.doi.org/10.17504/protocols.io.h7bb9in
null
null
TITLE: No Title AUTHORS: [DESCRIPTION] <p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/j4gdmie.png" data-src="https://s3.amazonaws.com/pr-journal/j4fdmie.png" data-ofn="cpib_root_analysis.png" /></p> [STEPS] ?. ?. ?. ?. ?. ?. ?.
[]
66,128
Protocol: A Systematic Review of Graves&#39; Disease and its Interconnection with MHC Class II
1
dx.doi.org/10.17504/protocols.io.cctqswmw
https://www.protocols.io/view/protocol-a-systematic-review-of-graves-39-disease-cctqswmw
Dylan Thibaut, Connor Sweeney, Shannon South, Mohamed Hussein
TITLE: Protocol: A Systematic Review of Graves&#39; Disease and its Interconnection with MHC Class II AUTHORS: Dylan Thibaut, Connor Sweeney, Shannon South, Mohamed Hussein [DESCRIPTION] Graves’ disease continues to be a problematic autoimmune thyroid condition debilitating patients with hyperthyroidism. HLAs play ...
["[Administrative Information] Title\n\"A Systematic Review of Graves' Disease and its Interconnection with MHC Class II\"\n\nRegistration \nprotocols.io", "[Introduction] Rationale\n Through meta-analysis, data from research can be extracted and evaluated to find specific HLA-DRB1 and HLA-DQ molecules associated w...
46,675
Flow Cytometer Fluorescence Voltration for FCMPASS
1
dx.doi.org/10.17504/protocols.io.brttm6nn
https://www.protocols.io/view/flow-cytometer-fluorescence-voltration-for-fcmpass-brttm6nn
Joshua A Welsh, Sean M Cook, Jennifer Jones
TITLE: Flow Cytometer Fluorescence Voltration for FCMPASS AUTHORS: Joshua A Welsh, Sean M Cook, Jennifer Jones [DESCRIPTION] Protocol to perform flow cytometer voltration to identify optimal detector settings for small particle analysis. Data acquired from this protocol are compatible with semi-automated analysis too...
["[Sample preparation] Vortex bottle on a high setting for 5 s.", "[Sample preparation] Pipette 500 µL of to two . Label one tube 'DPBS\" and the second tube 'Beads'.", "[Sample preparation] Add 3 drops of to the 'Beads' and vortex for r 5 s.", "[Cytometer Setup] Ensure cytometer is clean and that -Height and...
54,748
Quantitative PCR of Viral Abundance
4
dx.doi.org/10.17504/protocols.io.bzp4p5qw
https://www.protocols.io/view/quantitative-pcr-of-viral-abundance-bzp4p5qw
Bryan Yoo, Jessica Griffiths, Sarkis Mazmanian
TITLE: Quantitative PCR of Viral Abundance AUTHORS: Bryan Yoo, Jessica Griffiths, Sarkis Mazmanian [DESCRIPTION] Protocol for quantifying viral abundance by PCR used in Yoo et al 2021 [STEPS] 1. AAV-PHP.S:hSYN1-mNeonGreen was delivered systemically to wildtype mice at 6-8 weeks of age. 2. 3-4 weeks following viral...
["AAV-PHP.S:hSYN1-mNeonGreen was delivered systemically to wildtype mice at 6-8 weeks of age.", "3-4 weeks following viral infection, 1cm of proximal, medial, and distal small intestine (SI) and colon were harvested and flash frozen in TRIzol (ThermoFisher Scientific, Waltham, MA-Cat. No. 15596018) and RNA was extracte...
27,204
Distribution of aerophilous diatom communities associated with terrestrial green macroalgae in the South Shetland Islands, Maritime Antarctica - PROTOCOL
null
dx.doi.org/10.17504/protocols.io.6tcheiw
null
Juliana Ferreira da Silva, Maria Angélica Oliveira, Raylane Ribeiro Anunciação, Eduardo Pereira da Silva, Rodrigo Paidano Alves, Adriano Luis Schunemann, Filipe de Carvalho Victoria, Margéli Pereira de Albuquerque, Antonio Batista Pereira
TITLE: Distribution of aerophilous diatom communities associated with terrestrial green macroalgae in the South Shetland Islands, Maritime Antarctica - PROTOCOL AUTHORS: Juliana Ferreira da Silva, Maria Angélica Oliveira, Raylane Ribeiro Anunciação, Eduardo Pereira da Silva, Rodrigo Paidano Alves, Adriano Luis Schunem...
[]
76,894
Ambrecht et al. 2020: An optimized method for the extraction of ancient eukaryote DNA from marine sediments
4
dx.doi.org/10.17504/protocols.io.x54v9dw3qg3e/v1
https://www.protocols.io/view/ambrecht-et-al-2020-an-optimized-method-for-the-ex-cpb6vire
Linda Armbrecht
TITLE: Ambrecht et al. 2020: An optimized method for the extraction of ancient eukaryote DNA from marine sediments AUTHORS: Linda Armbrecht [DESCRIPTION] Four combinations of sedaDNA extraction treatments using marine sediments collected at a water depth of 104 m off Maria Island in Tasmania are compared. These metho...
["[Method 1: Bead-beating + spin column (DNeasy PowerLyzer PowerSoil Kit, Qiagen; “Kit”)] This technique was applied to 0.25 g of sediment subsamples stored at 4 °C, following the manufacturer's protocol with the some modifications:", "[Shotgun sequencing library preparation] We prepared four libraries as described in ...
37,028
Western blot analysis and immunoprecipitation assay
null
dx.doi.org/10.17504/protocols.io.bgecjtaw
https://www.protocols.io/view/western-blot-analysis-and-immunoprecipitation-assa-bgecjtaw
Marzia Ognibene
TITLE: Western blot analysis and immunoprecipitation assay AUTHORS: Marzia Ognibene [STEPS] ?. Cells are washed in cold PBS and lysed on ice in a buffer containing 1.6 mM NaH2PO4, 8.6 mM Na2HPO4, 1% Triton X-100, 0.1% SDS; 0.1% NaCl, 0.5% NaDoc, 2 mM AEBSF, 20 mg/mL each of aprotinin and leupeptin. ?. Cell lysates are...
["Cells are washed in cold PBS and lysed on ice in a buffer containing 1.6 mM NaH2PO4, 8.6 mM Na2HPO4, 1% Triton X-100, 0.1% SDS; 0.1% NaCl, 0.5% NaDoc, 2 mM AEBSF, 20 mg/mL each of aprotinin and leupeptin.", "Cell lysates are centrifuged in a microfuge at maximum speed for 10 min to eliminate debris.", "Protein conten...
28,743
Removal of gDNA from totalRNA
null
dx.doi.org/10.17504/protocols.io.8bfhsjn
null
iGEM Dusseldorf
TITLE: Removal of gDNA from totalRNA AUTHORS: iGEM Dusseldorf [DESCRIPTION] <div class = "text-blocks"><div class = "text-block">Removal of genomic DNA from RNA preparations</div></div> [STEPS] ?. [Digestion of DNA] pipet components into a RNase-free 1.5 ml tube (following order: H2O, buffer, total RNA, DNaseI) ?. [D...
["[Digestion of DNA]\npipet components into a RNase-free 1.5 ml tube (following order: H2O, buffer, total RNA, DNaseI)", "[Digestion of DNA]\nincubate reaction at 37°C", "[Extraction of DNaseI-digested RNA]\nadd 1 µl 50 mM EDTA and incubate at 65°C", "[Extraction of DNaseI-digested RNA]\nadd 1/10 Volume NaOAc and 3 Vol...
12,937
Electroporation of the protist Cafeteria roenbergensis cells using the Neon Transfection System
null
dx.doi.org/10.17504/protocols.io.qvhdw36
null
Sarah Duponchel, Monica Berjon-Otero, Matthias Fischer
TITLE: Electroporation of the protist Cafeteria roenbergensis cells using the Neon Transfection System AUTHORS: Sarah Duponchel, Monica Berjon-Otero, Matthias Fischer [DESCRIPTION] <div class = "text-blocks"></div> [STEPS] ?. [Cell preparation] Before the experiment (it can be the day before without affecting too muc...
["[Cell preparation]\nBefore the experiment (it can be the day before without affecting too much the cell viability)\n{\"blocks\":[{\"data\":[],\"depth\":0,\"entityRanges\":[],\"inlineStyleRanges\":[{\"length\":24,\"offset\":165,\"style\":\"italic\"}],\"key\":\"1ovn6\",\"text\":\"Heterotrophic flagellates require co-cu...
null
null
null
dx.doi.org/10.17504/protocols.io.k6tczen
null
null
TITLE: No Title AUTHORS: [GUIDELINES] <p>Overall Notes:</p> <ul> <li>Measure all regions twice (on separate days) <ul> <li>Use Pearson coefficient to determine how similar they are</li> <li>If Pearson coefficient is 0.9, then average them together</li> <li>If not, measure regions again and re-calculate the Pearson’s ...
[]
97,729
Open-source tools for Spatial Tissue Exploration Program (STEP) PhenoCycler-Fusion image analysis.
0
null
https://www.protocols.io/view/open-source-tools-for-spatial-tissue-exploration-p-dbn92mh6
Daniel H Geschwind, George Chen
TITLE: Open-source tools for Spatial Tissue Exploration Program (STEP) PhenoCycler-Fusion image analysis. AUTHORS: Daniel H Geschwind, George Chen [DESCRIPTION] Link to PhenoCycler User Guide to allow for multi-plex spatial proteomic detection on fresh frozen and FFPE tissue sections plus image analysis. [STEPS] SEC...
["[PhenoCycler Imaging Analysis protocol] Imaging Analysis protocol can be found here:\nThis protocol is working for us.", "[STEP protocol for PhenoCycler coverslip preparation] STEP protocol for PhenoCycler coverslip preparation can be found here: \nThis protocol is working for us."]
68,546
Hamster infestation with cercariae
4
null
https://www.protocols.io/view/hamster-infestation-with-cercariae-ce7athie
Frédéric D. Chevalier
TITLE: Hamster infestation with cercariae AUTHORS: Frédéric D. Chevalier [DESCRIPTION] This protocol describes the steps, materials and precautions needed for infecting hamsters with schistosome cercariae. Note that the use of vertebrates for research needs to be approved by the Institutional Animal Care and Use Commi...
["Shed cercariae from snails during 2h under light in TBRI water", "Discard the shedding water in a new beaker", "Shed cercariae from snails again during 2h under light in new TBRI water", "Transfer the water in a new dish", "Get the hamster", "Wet the hamster in jar containing hot water (~30°C). BE CAREFUL to keep alw...