id float64 1.55k 110k ⌀ | title stringlengths 1 256 ⌀ | template_id float64 0 6 ⌀ | doi stringlengths 39 49 ⌀ | url stringlengths 40 92 ⌀ | authors stringlengths 1 933 ⌀ | protocol_text stringlengths 34 1.08M | steps_list stringlengths 2 269k |
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null | null | null | dx.doi.org/10.17504/protocols.io.hv7b69n | null | null | TITLE: No Title
AUTHORS:
[GUIDELINES]
<strong>Materials: </strong><br />• Sterile PBS <br />• Anti-human CD3 Antibody <br /> - Clone UCHT1 (LEAF™ format, Cat. No. 300413/300414/300432; Ultra-LEAF™ format, Cat. No. 300437/300438)<br /> - Clone OKT3 (LEAF™ format, Cat. No. 317303/317304/317315; Ultra-LEAF™ ... | [] |
36,960 | sandwitch 1 | null | dx.doi.org/10.17504/protocols.io.bgb8jsrw | https://www.protocols.io/view/sandwitch-1-bgb8jsrw | null | TITLE: sandwitch 1
AUTHORS:
[STEPS] | [] |
47,711 | Isolation of live single cells from intestinal biopsy | 4 | dx.doi.org/10.17504/protocols.io.bst7nern | https://www.protocols.io/view/isolation-of-live-single-cells-from-intestinal-bio-bst7nern | Tatiana Karakasheva, Kathryn Hamilton | TITLE: Isolation of live single cells from intestinal biopsy
AUTHORS: Tatiana Karakasheva, Kathryn Hamilton
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes dissociation of a human intestinal biopsy tissue into single cells, followed by depletion of dead cells via annexin V MA... | ["[Before you begin]\nPrepare reagents: Cryo-wash (4.5mL PBS + 500μL FBS) Enzyme Mix (30μL Liberase TH stock + 30μL DNAse I stock + 3mL HBSS) Blocker Buffer (4% BSA in PBS: reconstitute 400mg BSA in 10mL PBS, filter-sterilize) Cell Reconstitution Buffer (2mL Advanced DMEM:F12 + 50μL Blocker Buffer + 20μL DNAse I) 1x M... |
28,828 | Golden Gate lvl 1/2 | null | dx.doi.org/10.17504/protocols.io.8d4hs8w | null | Vinca Seiler, René Inckemann | TITLE: Golden Gate lvl 1/2
AUTHORS: Vinca Seiler, René Inckemann
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Golden Gate reaction protocol for lvl 1/2</div></div>
[STEPS]
?. [Pipetting scheme for assembly reaction]
of each DNA insert. For an improved assembly efficiency, the amount of DNA inser... | ["[Pipetting scheme for assembly reaction]\nof each DNA insert. For an improved assembly efficiency, the amount of DNA inserts can optionally be normalized to equimolar concentrations (∼ each) or use of each insert (antibiotic resistance part should be diluted 1:10, -).\n0.5 µl\n20\n75 ng\n7.5 ng\n10 ng", "[Pipetting ... |
61,684 | Thawing of CT-2A cell line | 4 | null | https://www.protocols.io/view/thawing-of-ct-2a-cell-line-b8gurtww | Guido Krähenbühl | TITLE: Thawing of CT-2A cell line
AUTHORS: Guido Krähenbühl
[DESCRIPTION]
This protocol describes the thawing of mouse CT-2A glioma cells
[BEFORE_START]
Make sure to warm up media to 37 °C
[GUIDELINES]
GMO safety level 1
[STEPS]
1. Remove cryovial from liquid nitrogen storage and place on dry-ice for transfer t... | ["Remove cryovial from liquid nitrogen storage and place on dry-ice for transfer to cell culture lab.", "Remove cryovial from dry-ice and hold in a 37 °C waterbath until thawed\n \n\nDry the cryovial thoroughly, spray with 70% ethanol and transfer to cell culture hood", "Transfer contents of cryovial to a containin... |
41,276 | CDx autoXpress detection of COVID 19 | 4 | dx.doi.org/10.17504/protocols.io.bki4kugw | https://www.protocols.io/view/cdx-autoxpress-detection-of-covid-19-bki4kugw | Mickey Shah, Gary Niehaus | TITLE: CDx autoXpress detection of COVID 19
AUTHORS: Mickey Shah, Gary Niehaus
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The Crystal Diagnostics AutoXpress provides rapid detection of COVID 19 at 10 - 100 CFU per ul of clinical sample (nasal swab, sputum, or saliva). The patented Liquid Crysta... | ["[Automated]\nTransfer of test sample to 96 well plate. Each sample will be transferred to two wells, one for POS and one for NEG.\n600 µl", "[Automated]\nMix the magnetic 3um microspheres for POS and NEG using a pipette.", "[Automated]\nDispense of the magnetic 3um microspheres to each well.\n100 µl", "[Automated]\... |
45,022 | University of Helsinki and Natural Resources Institute Finland (Luke) protocol for DNA extraction and multiplex PCR genotyping of 17 microsatellites for pikeperch (Sander lucioperca L.). | 4 | dx.doi.org/10.17504/protocols.io.bp76mrre | https://www.protocols.io/view/university-of-helsinki-and-natural-resources-insti-bp76mrre | Jarmo Koskiniemi, Marja-Liisa Koljonen, Tuomas Leinonen | TITLE: University of Helsinki and Natural Resources Institute Finland (Luke) protocol for DNA extraction and multiplex PCR genotyping of 17 microsatellites for pikeperch (Sander lucioperca L.).
AUTHORS: Jarmo Koskiniemi, Marja-Liisa Koljonen, Tuomas Leinonen
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-... | ["DNA is extracted from dried scales or from fins or other tissues preserved in alcohol, frozen or fresh. The extractions are done using Qiagen DNeasy or DNAeasy 96 Blood & Tissue Kits with the kit manual’s ‘Animal Tissues’ protocols with a few modifications for the egg samples.", "Usually only 1 scale, or if they are ... |
72,235 | Protocol 5: Agrobacterium-mediated transformation of Spizellomyces punctatus (Sp) | 4 | dx.doi.org/10.17504/protocols.io.n92ldpp5xl5b/v1 | https://www.protocols.io/view/protocol-5-agrobacterium-mediated-transformation-o-cisjuecn | Sarah M Prostak, Edgar M Medina, Erik Kalinka, Lillian Fritz-Laylin | TITLE: Protocol 5: Agrobacterium-mediated transformation of Spizellomyces punctatus (Sp)
AUTHORS: Sarah M Prostak, Edgar M Medina, Erik Kalinka, Lillian Fritz-Laylin
[DESCRIPTION]
Once you have completed protocols 1-4, you are ready for transformation. This protocol is lengthy, requires accurate timing, and takes a mi... | ["[Growing Agro to the appropriate OD660] Dilute each overnight Agro culture to an OD660 = 0.15 using IM.", "[Growing Agro to the appropriate OD660] Grow the Agro at 28 °C and shaking at until OD660 = 0.6.", "[Harvesting Sp zoospores for transformation] Flood all wild type Sp plates with 1 mL of DS or IM for 60 min."... |
99,658 | Genotyping Wheat Grains to Identify Parental Donation of Alleles | 0 | dx.doi.org/10.17504/protocols.io.6qpvr84xplmk/v1 | https://www.protocols.io/view/genotyping-wheat-grains-to-identify-parental-donat-ddji24ke | Delfi Dorussen, Samuel Burrows, Giorgia Di Santolo, Philippa Borrill | TITLE: Genotyping Wheat Grains to Identify Parental Donation of Alleles
AUTHORS: Delfi Dorussen, Samuel Burrows, Giorgia Di Santolo, Philippa Borrill
[DESCRIPTION]
In wheat research, KASP (Kompetitive Allele Specific PCR) is widely used to determine the genotypes of progeny from large, segregating populations. However... | ["[Grain Surface Sterilisation] Make up the sterilisation solution: add 5 mL sodium hypochlorite solution to 100 mL sterile diH2O.", "[Grain Surface Sterilisation] Place the grains in a 15 mL Falcon tube and add 5 mL 70% ethanol. Vortex the tube for 10 seconds, leave for 5 minutes, and vortex again for 10 seconds.", "[... |
106,086 | LogP / LogD shake-flask method | 0 | dx.doi.org/10.17504/protocols.io.e6nvw14kdlmk/v1 | https://www.protocols.io/view/logp-logd-shake-flask-method-djue4nte | Yurii Kheylik | TITLE: LogP / LogD shake-flask method
AUTHORS: Yurii Kheylik
[DESCRIPTION]
Lipophilicity is possibly the most important physicochemical parameter for any potential drug candidate. Lipophilicity measurements are valuable for understanding how drugs are dissolved in plasma and other aqueous biological fluids. Lipophilic... | ["[Preparation of auxiliary solutions] 1 litre PBS: 0.01 M Phosphate buffer\n\nIn a bottle for reagents with a cap of 1L capacity, place:\n\ncontents of 1 L sachet with phosphate buffer,\n1 L of water, mix thoroughly, and filter through a membrane filter with a pore diameter of 0.45 μm.", "[Preparation of auxiliary sol... |
49,143 | Protocolo Experimento - Competição | 5 | null | https://www.protocols.io/view/protocolo-experimento-competi-o-bt8xnrxn | Fabiano Santos Conrado | TITLE: Protocolo Experimento - Competição
AUTHORS: Fabiano Santos Conrado
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">PiCOC</div><div class = "text-block"><ol style = "list-style-type: decimal;"></ol></div><div class = "text-block"> 2. Intervenção:<padrao de design gamificado 1></div><div clas... | ["[Pre-teste]\nO professor dará uma breve explicação sobre o experimento e solicitará que os alunos avaliem e preencham os formulários (Participação deverá ser voluntária)Formulário de Pré-teste (DFS-STAI-T) Formulário de Pré-Teste | Programação WEB 1 (3º Ano) 08/06/2021 15h-16h40Formulário de Pré-teste (DFS-STAI-T) F... |
36,100 | A protocol for rapid detection of the 2019 novel coronavirus SARS-CoV-2 using CRISPR diagnostics: SARS-CoV-2 DETECTR | null | dx.doi.org/10.17504/protocols.io.bfhcjj2w | https://www.protocols.io/view/a-protocol-for-rapid-detection-of-the-2019-novel-c-bfhcjj2w | Mammoth Biosciences, James P. Broughton, Wayne Deng, Clare L. Fasching, Jasmeet Singh, Charles Y. Chiu, Janice S. Chen | TITLE: A protocol for rapid detection of the 2019 novel coronavirus SARS-CoV-2 using CRISPR diagnostics: SARS-CoV-2 DETECTR
AUTHORS: Mammoth Biosciences, James P. Broughton, Wayne Deng, Clare L. Fasching, Jasmeet Singh, Charles Y. Chiu, Janice S. Chen
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">... | ["[Prepare nucleic acid sample and CRISPR reagents]\nExtract patient RNA following CDC recommendations.", "[Prepare nucleic acid sample and CRISPR reagents]\nPrepare LbCas12a RNP complexes for the samples to be tested. One complex for N-gene, E-gene, and RNase P gRNAs is needed for each sample. ABC1ReagentVolumeFinal ... |
59,253 | RNA stability in SPECTRA | 4 | null | https://www.protocols.io/view/rna-stability-in-spectra-b54vq8w6 | Venkata Subramanian Ramesan, Bhushan Toley | TITLE: RNA stability in SPECTRA
AUTHORS: Venkata Subramanian Ramesan, Bhushan Toley
[DESCRIPTION]
Protocol for storage of RNA in SPECTRA.
[STEPS]
SECTION: Source the RNA
1. Obtain any RNA (RNase P, SARS CoV-2, etc.)
SECTION: Source the RNA
2. Quantify the RNA using a Nanodrop or RT-PCR. Convert to copy number usi... | ["[Source the RNA] Obtain any RNA (RNase P, SARS CoV-2, etc.)", "[Source the RNA] Quantify the RNA using a Nanodrop or RT-PCR. Convert to copy number using the following tool.\n\nRNA copy number calculator", "[Apply RNA to the nylon flock] Add 10ul of 10^4 copies /ul RNA to the swab", "[Load the swab into the SPECTRA]... |
104,833 | Induction of arthritis in rats with mBSA | 0 | dx.doi.org/10.17504/protocols.io.81wgbz9d1gpk/v1 | https://www.protocols.io/view/induction-of-arthritis-in-rats-with-mbsa-dik94cz6 | Lizandre Keren Ramos da Silveira, Ana Paula P. Velosa, Sergio Catanozi, Marco Aurélio A. Pereira, Antonio dos Santos Filho, Fabio Luiz N. Marques, Daniele de Paula Faria, Caroline Cristiano Real, Sandra de M. Fernezlian, Amanda Flores Yanke, Zelita Aparecida de J. Queiroz, Vitória Elias Contini, Thays de Matos Lobo, So... | TITLE: Induction of arthritis in rats with mBSA
AUTHORS: Lizandre Keren Ramos da Silveira, Ana Paula P. Velosa, Sergio Catanozi, Marco Aurélio A. Pereira, Antonio dos Santos Filho, Fabio Luiz N. Marques, Daniele de Paula Faria, Caroline Cristiano Real, Sandra de M. Fernezlian, Amanda Flores Yanke, Zelita Aparecida de J... | ["[Materials and Methods] Experimental protocol\nTwenty-five three-month-old male Lewis rats with an average body mass of 360g were provided by the animal facility of our institution. All of the experimental procedures were approved by The Committee on Ethical Use of Laboratory Animals of the Faculty of Medicine at the... |
28,621 | MC38 and B16-F10 culture media | null | dx.doi.org/10.17504/protocols.io.77mhrk6 | null | Elinor Gottschalk, Bulent Arman Aksoy, Pinar Aksoy, Jeff Hammerbacher | TITLE: MC38 and B16-F10 culture media
AUTHORS: Elinor Gottschalk, Bulent Arman Aksoy, Pinar Aksoy, Jeff Hammerbacher
[STEPS]
?. [Mix components and sterile filter]
Working in the hood, remove 50 ml of DMEM from the 500 ml bottle (HyClone™ Dulbecco's High Glucose Modified Eagles Medium). To the remaining 450 ml DMEM ad... | ["[Mix components and sterile filter]\nWorking in the hood, remove 50 ml of DMEM from the 500 ml bottle (HyClone™ Dulbecco's High Glucose Modified Eagles Medium). To the remaining 450 ml DMEM add: 50 ml FBS (Fetal Plus) (10%) and 5 ml penicillin-streptomycin (1%).Sterile filter into a 500 ml 0.22 filter bottle. Store a... |
66,883 | Mass-Spectrometry analysis of ATP13A2 samples | 6 | dx.doi.org/10.17504/protocols.io.n92ldzdyxv5b/v1 | https://www.protocols.io/view/mass-spectrometry-analysis-of-atp13a2-samples-cdjbs4in | Sue Sim, eunyong_park | TITLE: Mass-Spectrometry analysis of ATP13A2 samples
AUTHORS: Sue Sim, eunyong_park
[DESCRIPTION]
Preparing mass-spectrometry samples to analyze polyamine content in purified ATP13A2 samples
[STEPS]
1. Pool purified ATP13A2 sample after SEC (sample should be in MS buffer after SEC)
2. Concentrate the protein to ~1... | ["Pool purified ATP13A2 sample after SEC (sample should be in MS buffer after SEC)", "Concentrate the protein to ~1.2 mg/mL using an Amicon Ultrafilter (cut-off 100kDa) at 4 °C", "Prepare standards of spermidine (Sigma-Aldrich) and spermine (Sigma-Aldrich) at a concentration of 10 µM in MS buffer", "Flash-freeze sample... |
42,666 | Nano-DESI Mass Spectrometry Imaging kidney characterization pipeline for tissues collected by Vanderbilt University | 1 | dx.doi.org/10.17504/protocols.io.bmwik7ce | https://www.protocols.io/view/nano-desi-mass-spectrometry-imaging-kidney-charact-bmwik7ce | yang1832 , Julia Laskin | TITLE: Nano-DESI Mass Spectrometry Imaging kidney characterization pipeline for tissues collected by Vanderbilt University
AUTHORS: yang1832 , Julia Laskin
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Our goal is to map the biological tissues with Nano-DESI Mass Spectrometry Imaging and build an ... | ["Collection of post-surgical tissue by Vanderbilt.Collection: dx.doi.org/10.17504/protocols.io.7gehjte", "Stabilize and freeze tissues by VanderbiltFreezing Tissue: dx.doi.org/10.17504/protocols.io.6wghfbw", "Intial Rapid Pathology Assessment of Kidney Tissue by Vanderbilt.Staining: dx.doi.org/10.17504/protocols.io.... |
72,270 | Quantification-Protocol-miRNA-SCALONMC | 6 | null | https://www.protocols.io/view/quantification-protocol-mirna-scalonmc-citnueme | Marcela Scalon, Christine Souza Martins, Gabriel Ginani Ferreira, Franciele Schlemmer, Ricardo Titze de Almeida, Giane Regina Paludo | TITLE: Quantification-Protocol-miRNA-SCALONMC
AUTHORS: Marcela Scalon, Christine Souza Martins, Gabriel Ginani Ferreira, Franciele Schlemmer, Ricardo Titze de Almeida, Giane Regina Paludo
[DESCRIPTION]
This protocol is intended as a guideline for the quantification of total miRNAs, from serum and plasma samples, using... | ["Prepare the Qubit® working solution by diluting the Qubit® microRNA reagent 1:200 in Qubit® microRNA buffer as follows:\n\n1µl of Qubit® microRNA reagent for each sample/standard\n199µl of Qubit® microRNA buffer for each sample/standard\n\nNote: Use a clean plastic tube each time you make Qubit® working solution. Do ... |
15,804 | Protocol of sampling of feces in preterm infants | null | dx.doi.org/10.17504/protocols.io.tn4emgw | null | Yuan Zhenya | TITLE: Protocol of sampling of feces in preterm infants
AUTHORS: Yuan Zhenya
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol refer to a study having the aim to study on the variation of gut microbiota in preterm infants in the process of feeding intolerance after birth.
In this study,... | ["[Fecal sampling]\nFecal samples were collected by disposable sterile swabs from the diapers of preterm infants, transferred to sterile tubes, and stored at -80℃ for the subsequent DNA extraction.", "[DNA extraction]\nThe bacterial genomic DNA from samples was extracted by E.Z.N.A.® Soil DNA Kit (Omega Bio-tek, Norcro... |
54,284 | Multi-layer Hybrid Classification Model of COVID-19 Chest X-ray Images | 5 | dx.doi.org/10.17504/protocols.io.by9kpz4w | https://www.protocols.io/view/multi-layer-hybrid-classification-model-of-covid-1-by9kpz4w | Thanakorn Poomkur | TITLE: Multi-layer Hybrid Classification Model of COVID-19 Chest X-ray Images
AUTHORS: Thanakorn Poomkur
[DESCRIPTION]
The coronavirus disease of 2019(COVID-19) has been declared a pandemic and has raised worldwide concern. Lung inflammation and respiratory failure are commonly observed in moderate-to-severe cases. R... | [] |
null | null | null | dx.doi.org/10.17504/protocols.io.f28bqhw | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p><em>Monitor </em><em>fluorescence of cyanobacterial cultures over time using a 96-well plate format. </em></p>
<p> </p>
[STEPS]
?.
?.
?.
?. | [] |
null | null | null | dx.doi.org/10.17504/protocols.io.pw8dphw | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>For investigate the presence of YFV genome by quantitative PCR assays in the hydrolysis probe detection format. The target is the 5’-noncoding region (5’-NC) of the YFV genome (Domingo et al., 2012).</p>
[STEPS]
?.
?.
?.
?.
?.
?.
?.
?.
?.
?.
?.
?.
?. | [] |
46,655 | Neuromuscular Junction Immunostaining | 1 | dx.doi.org/10.17504/protocols.io.brs7m6hn | https://www.protocols.io/view/neuromuscular-junction-immunostaining-brs7m6hn | Jake Willows, Kristy Townsend, Magdalena Blaszkiewicz | TITLE: Neuromuscular Junction Immunostaining
AUTHORS: Jake Willows, Kristy Townsend, Magdalena Blaszkiewicz
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">A series of protocols to fluorescently label neuromuscular junctions in mice for qualitative and quantitative studies.</div></div>
[STEPS]
?. ... | ["See attachments under Abstract for PDF copies of protocols."] |
23,832 | RbCl Uber-Competent Cells | null | dx.doi.org/10.17504/protocols.io.3hygj7w | null | Sricharan Kadimi | TITLE: RbCl Uber-Competent Cells
AUTHORS: Sricharan Kadimi
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">A protocol kindly provided to our team by the advisor to the 2018 MIT iGEM team (Dr. Brian Teague). It is modified slightly from that version (made by Stefan Maas in 1997) to fit our needs. Ste... | ["[Day 0]\nTake a tube of comercially competent cells and put it on ice (Thermo-Fisher TOP10 preferred, we think). Make sure to use a tube of comercially competent cells to start, or you may have poorer results. Work near a flame to ensure sterility.Using a sterile pipet tip or a flamed and cooled inoculation loop, tak... |
null | null | null | dx.doi.org/10.17504/protocols.io.h6gb9bw | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>Protocol for protein extraction, alkylation, and digestion to obtain LC/MS (liquid chromatography-mass spectrometry) data of calculated protein signal intensities in HEK-293 cells to monitor them.</p>
[BEFORE_START]
<p>Prepare a Krebs-Ringer-Buffer: </p>
<ul>
<li>154 mM NaCl... | [] |
null | null | null | dx.doi.org/10.17504/protocols.io.habb2an | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p><span style="font-weight: 400;">Electroporation of V. natriegens cells is a rapid 2-hour protocol, from culture inoculation to plating. Each transformation produces >10^5 CFU / µg plasmid DNA. Each transformation requires 10mL of LB3 (high-salt) media, corresponding to 50u... | [] |
null | null | null | dx.doi.org/10.17504/protocols.io.jx4cpqw | null | null | TITLE: No Title
AUTHORS:
[STEPS]
?. | [] |
100,000 | Zeiss AxioImager Multi Channel 20X Image Capture | 1 | dx.doi.org/10.17504/protocols.io.bp2l6n85rgqe/v3 | https://www.protocols.io/view/zeiss-axioimager-multi-channel-20x-image-capture-ddv8269w | Allen Institute for Brain Science | TITLE: Zeiss AxioImager Multi Channel 20X Image Capture
AUTHORS: Allen Institute for Brain Science
[DESCRIPTION]
This protocol describes the capture of 20X multi-channel images using human and mouse tissue.
Note: Research reported in this publication was supported by the National Institute Of Mental Health of the Nat... | [] |
28,312 | Metatranscriptomic screening for genes of interest | null | dx.doi.org/10.17504/protocols.io.7vyhn7w | null | Helena Pound, Steven Wilhelm | TITLE: Metatranscriptomic screening for genes of interest
AUTHORS: Helena Pound, Steven Wilhelm
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Find genes of interest in assembled metatranscriptomic library using blast.</div></div>
[STEPS]
?. [Establishing Candidates]
Blast assembly against referen... | ["[Establishing Candidates]\nBlast assembly against reference database you just created - e-value can be changed to your preference - final.txt will produce list of contigs from assembly that had blast hits, to be referred to as \"candidates\"", "[Establishing Candidates]", "[Establishing Candidates]", "[Establis... |
43,855 | Cell Lysis and Sonication | 4 | dx.doi.org/10.17504/protocols.io.bn3pmgmn | https://www.protocols.io/view/cell-lysis-and-sonication-bn3pmgmn | Vasso Makrantoni, Daniel Robertson, Adele L. Marston | TITLE: Cell Lysis and Sonication
AUTHORS: Vasso Makrantoni, Daniel Robertson, Adele L. Marston
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">A plethora of biological processes like gene transcription, DNA replication, DNA recombination, and chromosome segregation are mediated through protein–DNA i... | ["[Cell Lysis and Sonication]\nThaw cells . Add 1 volume (–) supplemented with 0.5% SDS, and protease inhibitors (Roche Complete EDTA-free tablet).\non ice\n0.3 mL\n[ice-cold 1× FA lysis buffer]\n[PMSF]\nFor calibrated ChIP-seq use a 2:1 ratio of S. cerevisiae to S. pombe cells, as measured by OD600, mix pellets of di... |
89,600 | Magnetic resonance imaging (MRI) | 1 | dx.doi.org/10.17504/protocols.io.bp2l6x3mdlqe/v1 | https://www.protocols.io/view/magnetic-resonance-imaging-mri-c3q8ymzw | Núria Peñuelas | TITLE: Magnetic resonance imaging (MRI)
AUTHORS: Núria Peñuelas
[DESCRIPTION]
Magnetic resonance imaging (MRI) of mouse brain
[STEPS]
1. Prepare the equipment: 7T Bruker BioSpec 70/30USR scanner (Bruker BioSpin GmbH, Ettlingen, Germany) equipped with a mini-imaging gradient set (400mT/m) and using a 72-mm inner diame... | ["Prepare the equipment: 7T Bruker BioSpec 70/30USR scanner (Bruker BioSpin GmbH, Ettlingen, Germany) equipped with a mini-imaging gradient set (400mT/m) and using a 72-mm inner diameter linear volume coil as a transmitter and a dedicated mouse brain surface coil as a receiver.", "Prepare the software: Acquire and proc... |
81,676 | AAV capsid library production | 4 | dx.doi.org/10.17504/protocols.io.5jyl8jyz9g2w/v1 | https://www.protocols.io/view/aav-capsid-library-production-ctzkwp4w | Miguel Chuapoco | TITLE: AAV capsid library production
AUTHORS: Miguel Chuapoco
[DESCRIPTION]
We recently developed adeno-associated virus (AAV) capsids to facilitate efficient and noninvasive gene transfer to the central and peripheral nervous systems. However, a detailed protocol for generating and systemically delivering novel AAV v... | ["[Reagent setup: Plasmid DNA] Grow bacterial stocks in LB or Plasmid+ media containing the appropriate selective antibiotic; refer\nto the Addgene catalog for suggested growth conditions. Use a large-scale endotoxin-free plasmid purification kit to isolate plasmids; elute plasmid DNA with the supplied Tris-EDTA (TE) b... |
65,393 | GrownMD Male Enhancement Gummies Shocking Price Update | 1 | dx.doi.org/10.17504/protocols.io.ewov1nk5kgr2/v1 | https://www.protocols.io/view/grownmd-male-enhancement-gummies-shocking-price-up-cb4rsqv6 | purecalmot | TITLE: GrownMD Male Enhancement Gummies Shocking Price Update
AUTHORS: purecalmot
[DESCRIPTION]
On the off chance that you are not a substance with your sexual show, you have gone to the ideal spot. In a general sense 65% of men ought to be taller under the midriff. Additionally, 70% of men need to have the choice t... | [] |
19,802 | Leaf surface acylsugar extraction and LC-MS profiling - v1.0 | null | dx.doi.org/10.17504/protocols.io.xj2fkqe | null | Yann-Ru Lou, Bryan Leong | TITLE: Leaf surface acylsugar extraction and LC-MS profiling - v1.0
AUTHORS: Yann-Ru Lou, Bryan Leong
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><ul style = "list-style-type:disc;"></ul></div><div class = "text-block"><ul style = "list-style-type:disc;"><li style = "counter-reset:ol0;list-style... | ["[Reagent preparation]\nPrepare the Acylsugar Extraction Solution (3:3:2 Acetonitrile:Isopropanol:water, 0.1% formic acid, 1 µM telmisartan (internal standard))To prepare Acylsugar Extraction Solution, combine:AcetonitrileIsopropanolddH2OFormic Acid (88%) (0.1% final conc.)1 mM Telmisartan (dissolved in 100% DMSO; fin... |
92,330 | DQ-BSA and BMV109 microscopy | 4 | dx.doi.org/10.17504/protocols.io.261gedk5ov47/v1 | https://www.protocols.io/view/dq-bsa-and-bmv109-microscopy-c6eizbce | Rebecca Wallings | TITLE: DQ-BSA and BMV109 microscopy
AUTHORS: Rebecca Wallings
[DESCRIPTION]
DQ-BSA and BMV109 microscopy for peritoneal macrophages
[STEPS]
1. BMV109 Pan Cathepsin probe (Vergent Bioscience) and DQ red BSA (Invitrogen) were added to each well at a final concentration of 1µM and 10µG/mL, respectively, and cells were i... | ["BMV109 Pan Cathepsin probe (Vergent Bioscience) and DQ red BSA (Invitrogen) were added to each well at a final concentration of 1µM and 10µG/mL, respectively, and cells were incubated at\n37°C for 1-hour. \n\nCells were washed 3 x DPBS +/+ and fixed for 10 minutes at room temperature in Invitrogen‱ eBioscience‱ Intra... |
76,185 | Adherent Cell Culture (generic) | 4 | dx.doi.org/10.17504/protocols.io.dm6gpjx3dgzp/v1 | https://www.protocols.io/view/adherent-cell-culture-generic-cnmzvc76 | Gwen Phung, Philippa R Kennedy | TITLE: Adherent Cell Culture (generic)
AUTHORS: Gwen Phung, Philippa R Kennedy
[DESCRIPTION]
Standard procedure for culturing adherent cell lines. Follow these instructions and cell type-specific instructions on the Cell line information protocol
[BEFORE_START]
Warm complete media in 37oC water bath for at least 20 m... | ["Remove spent media from flask.", "Add 5-10 mL of PBS. Gently swirl flask to rinse cells then remove PBS.\n - this removes serum proteins that might interfere with trypsin", "Add trypsin (0.05%, Gibco Cat. No. 25300054)\n1 mL for T25\n2.5 mL for T75\n5 mL for T150", "Incubate flask at 37oC for 5-10 min. \n- check t... |
102,361 | Quantification of varietal aroma compounds, oak markers, and Strecker aldehydes in wines by SPE and gas chromatography–tandem mass spectrometrytry | 0 | dx.doi.org/10.17504/protocols.io.dm6gpzd18lzp/v1 | https://www.protocols.io/view/quantification-of-varietal-aroma-compounds-oak-mar-df7z3rp6 | Cécile Thibon, Emilie Suhas, Alexandre Pons | TITLE: Quantification of varietal aroma compounds, oak markers, and Strecker aldehydes in wines by SPE and gas chromatography–tandem mass spectrometrytry
AUTHORS: Cécile Thibon, Emilie Suhas, Alexandre Pons
[DESCRIPTION]
This protocol outlines the conditions for the assaying volatile impact compounds in white and red... | ["[Extraction of volatile compounds] The SPE procedure was automated with a Gilson GX-274 ASPEC solid-phase extraction system (Villiers-Le-Bel, France). HR-X CHROMABOND (500 mg, 6mL, Nacherey-Nagel, France) was first activated with methanol (7 mL, 6 mL/min), washed twice with ultrapure water/ethanol (90/10, v/v; 2 mL, ... |
53,690 | Lipidomic analysis of tissue culture cells, tissues, and purified organelles | 4 | dx.doi.org/10.17504/protocols.io.byn2pvge | https://www.protocols.io/view/lipidomic-analysis-of-tissue-culture-cells-tissues-byn2pvge | Annie Jen, Lia Serrano, Katherine A. Overmyer, Joshua J. Coon | TITLE: Lipidomic analysis of tissue culture cells, tissues, and purified organelles
AUTHORS: Annie Jen, Lia Serrano, Katherine A. Overmyer, Joshua J. Coon
[DESCRIPTION]
Lipids are a class of molecules that have roles in energy storage, plasma membrane integrity, and signaling events. To gain more understanding of ... | ["[Sample Preparation Procedures] Plasma Preparation", "[Sample Preparation Procedures] Remove plasma samples from frozen conditions, immediately place on ice, and allow to thaw.\n\nTransfer 10 µL into a 1.5 mL microfuge tube using a pipette. Ensure to maintain samples on ice for the remainder of the extraction.", "[Sa... |
77,334 | Potassium phosphate buffer (0.9 M, pH 7.7) | 4 | null | https://www.protocols.io/view/potassium-phosphate-buffer-0-9-m-ph-7-7-cprwvm7e | Andreas Sagen | TITLE: Potassium phosphate buffer (0.9 M, pH 7.7)
AUTHORS: Andreas Sagen
[DESCRIPTION]
Potassium phosphate buffers are buffers with a buffering capacity between 5.8 and 8.0, where it is possible to adjust the pH and buffer strength with different ratios of Sodium phosphate monobasic and Sodium phosphate dibasic.
[STE... | ["[500 mL Phosphate buffer (0.9 M, pH 7.7)] Fill a sterile 500 mL bottle with 400 mL distilled water", "[500 mL Phosphate buffer (0.9 M, pH 7.7)] Measure 62.705 g Potassium phosphate dibasic and 11.568 g Potassium phosphate monobasic\n\nMaterials:", "[500 mL Phosphate buffer (0.9 M, pH 7.7)] Add dry ingredients and mix... |
81,379 | 10x Protocols: Tissue Fixation & Dissociation for Chromium Fixed RNA Profiling -- University of Minnesota TMCs (CG000553 Rev A) | 1 | dx.doi.org/10.17504/protocols.io.q26g7y2pkgwz/v1 | https://www.protocols.click/view/10x-protocols-tissue-fixation-amp-dissociation-for-ctqbwmsn | 10x Genomics | TITLE: 10x Protocols: Tissue Fixation & Dissociation for Chromium Fixed RNA Profiling -- University of Minnesota TMCs (CG000553 Rev A)
AUTHORS: 10x Genomics
[DESCRIPTION]
10x Genomics protocol where you fix the whole tissue and then dissociate it in preparation for Chromium Single Cell Expression flex
Protocol I... | [] |
53,569 | Endosomal and lysosomal immunoprecipitation for proteomics, lipidomics, and TEM | 4 | dx.doi.org/10.17504/protocols.io.byi9puh6 | https://www.protocols.io/view/endosomal-and-lysosomal-immunoprecipitation-for-pr-byi9puh6 | Hankum Park, Frances V Hundley, J. Wade Harper | TITLE: Endosomal and lysosomal immunoprecipitation for proteomics, lipidomics, and TEM
AUTHORS: Hankum Park, Frances V Hundley, J. Wade Harper
[DESCRIPTION]
Previous studies have developed methods for isolation of lysosomes, mitochondria, and peroxisomes from non-denaturing extracts. Here we describe an approach for p... | ["[Lysosomal immunoprecipitation (Lyso-IP) for organelle proteomics] Seed 293 cells or 293EL cells expressing TMEM192-HA and FLAG-EEA1 in 15-cm dishes, with one dish per replicate. Creation of the 293EL cells is described in protocol dx.doi.org/10.17504/protocols.io.byi7puhn.", "[Lysosomal immunoprecipitation (Lyso-IP)... |
null | null | null | dx.doi.org/10.17504/protocols.io.icrcav6 | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>Please contact Dr. Steven Wilhelm (wilhelm@utk.edu) for additional information regarding this protocol.</p>
<p> </p>
<p>Modified from Rioux, C., Jordan, D., & Rattray, J. B. (1983). Colorimetric determination of catechol siderophores in microbial cultures. <em>Analytical ... | [] |
59,274 | Inserting RFID into rats | 4 | dx.doi.org/10.17504/protocols.io.rm7vzyxzrlx1/v1 | https://www.protocols.io/view/inserting-rfid-into-rats-b55iq84e | a.beeson , Leah Solsberg-Woods, Gabriel J Barrero, Oksana Polesskaya, Abraham Palmer | TITLE: Inserting RFID into rats
AUTHORS: a.beeson , Leah Solsberg-Woods, Gabriel J Barrero, Oksana Polesskaya, Abraham Palmer
[DESCRIPTION]
Insert RFID chips into rats
[GUIDELINES]
Save all old cage cards for verification of DOB. Weaned Pups will be housed in Coleman B113 for at least 2 days before transported t... | ["Carefully install one RFID into the clean needle", "scruff rat skin on the back of neck with fingers", "Inset the sharp tips of the needle under the skin, target the back of the neck, close to the head.", "Release the RFID by pushing the plunger of the syringe", "Scan and make sure the RFID is under the skin", "uploa... |
72,147 | Light-Seq Cell Barcoding | 4 | dx.doi.org/10.17504/protocols.io.eq2ly77jplx9/v1 | https://www.protocols.io/view/light-seq-cell-barcoding-ciptudnn | Jocelyn Y. Kishi, Ninning Liu, Emma R. West, Kuanwei Sheng, Jack J. Jordanides, Matthew Serrata, Constance L. Cepko, Sinem K. Saka, Peng Yin | TITLE: Light-Seq Cell Barcoding
AUTHORS: Jocelyn Y. Kishi, Ninning Liu, Emma R. West, Kuanwei Sheng, Jack J. Jordanides, Matthew Serrata, Constance L. Cepko, Sinem K. Saka, Peng Yin
[DESCRIPTION]
We present Light-Seq, an approach for multiplexed spatial indexing of intact biological samples using light-directed DNA ba... | ["[Protocol Overview] This protocol is for selective barcoding of cells seeded onto an Ibidi 18-well µ-slide for our Light-Seq paper. https://www.nature.com/articles/s41592-022-01604-1\n\nA separate protocol can be found for selective barcoding of tissue samples. \nhttps://www.protocols.io/view/light-seq-x54v9jno4g3e/v... |
93,405 | Visualization of LRRK2 filaments in 293T cells | 4 | dx.doi.org/10.17504/protocols.io.8epv5xpz4g1b/v1 | https://www.protocols.io/view/visualization-of-lrrk2-filaments-in-293t-cells-c7f5zjq6 | Eva Karasmanis, Kathryn S Hatch | TITLE: Visualization of LRRK2 filaments in 293T cells
AUTHORS: Eva Karasmanis, Kathryn S Hatch
[DESCRIPTION]
Visualization of LRRK2 filaments in 293T cells
GOAL: Express GFP-LRRK2 with or without DARPin E11 and quantify the percentage of cells with LRRK2 filaments in the presence and absence of MLi-2 in 293T cells.
... | ["[Day 1: fibronectin coating and cell plating] Visualization of LRRK2 filaments in 293T cells\n \nGOAL: Express GFP-LRRK2 with or without DARPin E11 and quantify the percentage of cells with LRRK2 filaments in the presence and absence of MLi-2 in 293T cells.\n\nConstructs needed: \n\n1) CMV-8xHISDaprin C12-FLAG\n2) CM... |
37,744 | Magnetic bead-based TREG-TEFF cell isolation from PBMC with Miltenyi CD4+CD25+ Regulatory T cell Isolation Kit | 1 | null | https://www.protocols.io/view/magnetic-bead-based-treg-teff-cell-isolation-from-bg4qjyvw | Marco Cosentino, Elisa Storelli, Alessandra Luini, Emanuela Rasini, Massimiliano Legnaro, Marco Ferrari, Franca Marino | TITLE: Magnetic bead-based TREG-TEFF cell isolation from PBMC with Miltenyi CD4+CD25+ Regulatory T cell Isolation Kit
AUTHORS: Marco Cosentino, Elisa Storelli, Alessandra Luini, Emanuela Rasini, Massimiliano Legnaro, Marco Ferrari, Franca Marino
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span ... | ["Isolate PBMCs according to the standard protocol \"Separation and purification of human PBMC from BUFFY COAT\" or \"Separation and purification of human PBMC from FRESH BLOOD\"", "OPTIONAL STEPSorting of TREG is quite long procedure. Especially in clinical studies with whole blood of enrolled subject, it is possible ... |
71,858 | Rapid sequencing gDNA Barcoding RBK2004 | 1 | null | https://www.protocols.io/view/rapid-sequencing-gdna-barcoding-rbk2004-ciesubee | Carlos Goller, Carly Sjogren | TITLE: Rapid sequencing gDNA Barcoding RBK2004
AUTHORS: Carlos Goller, Carly Sjogren
[DESCRIPTION]
This protocol has been adapted from Oxford Nanopore Technologies.
You have isolated, purified, and quantified high-molecular weight DNA from your bacterial isolate. You have also worked with your bacterium and grown in... | ["[Library Preparation] Thaw kit components at Room temperature , spin down briefly using a microfuge and mix by pipetting as indicated by the table below: \n\nFragmentation Mix RB01-12: not frozen, briefly spin down, mix well by pipetting \nRapid Adapter (RAP): not frozen, briefly spin down, mix well by pipetting \nSe... |
null | null | null | dx.doi.org/10.17504/protocols.io.unbevan | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
Illumina® short-read DNA sequencing has become an integral tool in biology for genome-wide studies. Offering accurate base-pair resolution at the most competitive price, the technology has become widespread. However, the generation of multiplexed DNA libraries remains costly and... | ["[DNA quantification] Ensure each well in the DNA plate is approximately below 20 ng/µL. For instance, quantify a few samples on a NanoDrop (ThermoFisher Scientific) and dilute the plate with nuclease-free water.", "[DNA quantification] {\"blocks\":[{\"key\":\"es0em\",\"text\":\"Create a Quant-iTTM master-mix by mixin... |
91,088 | Neuromelanin quantification in stained brain slices of the Locus coeruleus | 5 | dx.doi.org/10.17504/protocols.io.5qpvo32y9v4o/v1 | https://www.protocols.io/view/neuromelanin-quantification-in-stained-brain-slice-c47qyzmw | Csilla Novák, Maria P. Contreras, Matthias Prigge | TITLE: Neuromelanin quantification in stained brain slices of the Locus coeruleus
AUTHORS: Csilla Novák, Maria P. Contreras, Matthias Prigge
[DESCRIPTION]
This protocol describes an automatizable pipeline for cell counting based on fluorescent stainings with the help of CellPose. Additionally, it describes the entire... | ["[Image optimization and cropping] Start with images of 35μm thick slices of one z-plane and merged channels. In case of several z-planes, create a Maximum Intensity or a Sum Slices projection in ImageJ/Fiji.", "[Image optimization and cropping] Crop the area that you would like to analyze.", "[Image optimization and ... |
40,607 | Vezina Lab DIG-UTP Riboprobe Synthesis | 1 | null | https://www.protocols.io/view/vezina-lab-dig-utp-riboprobe-synthesis-bjv7kn9n | Chad Vezina | TITLE: Vezina Lab DIG-UTP Riboprobe Synthesis
AUTHORS: Chad Vezina
[STEPS]
?. [Design of Polymerase Chain Reaction (PCR) Primers]
Extract cDNA sequence from Entrez Gene (http://www.ncbi.nlm.nih.gov/sites/entrez).
?. [Design of Polymerase Chain Reaction (PCR) Primers]
Use Primer3 program (http://frodo.wi.mit.edu/primer... | ["[Design of Polymerase Chain Reaction (PCR) Primers]\nExtract cDNA sequence from Entrez Gene (http://www.ncbi.nlm.nih.gov/sites/entrez).", "[Design of Polymerase Chain Reaction (PCR) Primers]\nUse Primer3 program (http://frodo.wi.mit.edu/primer3/) to design PCR primers against the 3’-region of the cDNA sequence. The ... |
53,741 | Freequenza Box - Production and Assembly | 1 | dx.doi.org/10.17504/protocols.io.byqmpvu6 | https://www.protocols.io/view/freequenza-box-production-and-assembly-byqmpvu6 | Matthew Meyer, Katarzyna Dobaczewska, Zbigniew Mikulski | TITLE: Freequenza Box - Production and Assembly
AUTHORS: Matthew Meyer, Katarzyna Dobaczewska, Zbigniew Mikulski
[DESCRIPTION]
A histology aid for slide immunostaining. Improves staining consistency, reduces reagent use, and saves time. Works with Shandon Plastic Coverplates. For technical specifications, check out... | ["[3D-Printing] Navigate to the required 3D-printable files from the NIH 3D Print Exchange", "[3D-Printing] Select the desired model type from the below options and download the file in the \"Other Files\" section\n2x1 Box.zip - (2 slide capacity)\n3x2 Box.zip - (6 slide capacity)\n5x3 Box.zip - (15 slide capacity)\n7x... |
68,725 | Workflow for proteomic analysis of purified lysosomes in cells lacking GRN | 1 | dx.doi.org/10.17504/protocols.io.ewov14b7kvr2/v3 | https://www.protocols.io/view/workflow-for-proteomic-analysis-of-purified-lysoso-cfcvtiw6 | Sharan Swarup, J. Wade Harper | TITLE: Workflow for proteomic analysis of purified lysosomes in cells lacking GRN
AUTHORS: Sharan Swarup, J. Wade Harper
[DESCRIPTION]
Lysosomes are a major degradative organelle within eukaryotic cells. Previous work has developed a method wherein the TMEM192 protein is tagged on its C-terminus with an epitope tag i... | ["[Cell culture] Grow the appropriate cells (e.g. HeLa with or without the GRN gene created by gene editing, see DOI: dx.doi.org/10.17504/protocols.io.4r3l2oxqqv1y/v1) expressing TMEM192-3xHA in DMEM containing 10% FBS", "[Lyso-IP] All buffers were supplemented with protease and phosphatase inhibitors (Roche).", "[Lyso... |
49,478 | Bacterial Isolation of Mycobacterium avium | 1 | dx.doi.org/10.17504/protocols.io.bujenuje | https://www.protocols.io/view/bacterial-isolation-of-mycobacterium-avium-bujenuje | Tetsuya Komatsu, Kenji Ohya, Justice Opare Odoi, Shota Suganuma, Kotaro Sawai | TITLE: Bacterial Isolation of Mycobacterium avium
AUTHORS: Tetsuya Komatsu, Kenji Ohya, Justice Opare Odoi, Shota Suganuma, Kotaro Sawai
[DESCRIPTION]
Mycobacterium avium subsp. hominissuis (MAH) is one of the most important agents causing non-tuberculosis mycobacterial infection in human and pigs. Genome analysis o... | ["Cut out 20 mg of the caseous necrotic area and its surroundings from lymph nodes or organs and put them in a screw tube.", "Put some 5 mm stainless steel spheres and 400 µl of 2% NaOH in the screw tube.", "Set the tube in a tabletop crusher and process it at 3,200 rpm for 30 seconds.", "Add 400 µl of 0.5% N-Acetyl-L ... |
63,397 | Vissentials Max BHB Canada:-Uses, Side Effects, and More | 3 | dx.doi.org/10.17504/protocols.io.j8nlkk256l5r/v1 | https://www.protocols.io/view/vissentials-max-bhb-canada-uses-side-effects-and-m-b96dr9a6 | Max | TITLE: Vissentials Max BHB Canada:-Uses, Side Effects, and More
AUTHORS: Max
[DESCRIPTION]
Vissentials Max BHB Reviews, Canada: Obesity issues are fatal, and if you have an obese body, it may provide you multiple negative effects if you do not get rid of it in time. If you have an obese body, you will know the strug... | [] |
21,954 | MPE-seq | null | dx.doi.org/10.17504/protocols.io.zpaf5ie | null | Ben Fair | TITLE: MPE-seq
AUTHORS: Ben Fair
[STEPS]
?. [RNA fragmentation]
*Fragmentation may eliminate size biases against large primer extensions which are inefficiently PCR amplified and sequenced, but may be unnecessary depending on the application. I usually prefer not to fragment... If you want to do fragmentation, I recom... | ["[RNA fragmentation]\n*Fragmentation may eliminate size biases against large primer extensions which are inefficiently PCR amplified and sequenced, but may be unnecessary depending on the application. I usually prefer not to fragment... If you want to do fragmentation, I recommend fragmenting 15 to 40ug RNA by assembl... |
37,814 | High quality DNA extraction from very small individual insects | 4 | dx.doi.org/10.17504/protocols.io.bg6wjzfe | https://www.protocols.io/view/high-quality-dna-extraction-from-very-small-indivi-bg6wjzfe | Sam Mugford, Roland Wouters, Thomas C Mathers, Saskia Hogenhout | TITLE: High quality DNA extraction from very small individual insects
AUTHORS: Sam Mugford, Roland Wouters, Thomas C Mathers, Saskia Hogenhout
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Genomic studies of natural populations frequently benefit by having data from single individuals. This approa... | ["Grind a single frozen aphid in a 1.5ml Eppendorf tube, with a plastic pestle precooled in liquid nitrogen. Ensure that tissue remains frozen throughout by working with the Eppendorf tube sitting in a box of dry ice.", "Remove the tube from the dry ice and immediately add 100 µl CTAB buffer. Leave the plastic pestle i... |
93,347 | Histology, Immunohistochemistry and Imaging | 1 | dx.doi.org/10.17504/protocols.io.eq2lyjbdelx9/v1 | https://www.protocols.io/view/histology-immunohistochemistry-and-imaging-c7ebzjan | Jonathan Tang | TITLE: Histology, Immunohistochemistry and Imaging
AUTHORS: Jonathan Tang
[DESCRIPTION]
Sample preparation, histology, immunohistochemistry (IHC) and imaging of brain sections in Tang et al 2023.
[BEFORE_START]
Process begins after behavioral sessions were completed.
[STEPS]
SECTION: Sample Preparation
1. Deeply ... | ["[Sample Preparation] Deeply anesthetized mouse with isoflurane.", "[Sample Preparation] Perfuse transcardially in PBS and then 4% PFA/PBS.", "[Sample Preparation] Dissect brains with skulls attached and submerged in 4% PFA in PBS at 4 degrees Celsius overnight", "[Sample Preparation] Next day-brains were rinsed 3 tim... |
86,810 | CAMbank: cfDNA BCT Field Processing v1 | 4 | null | https://www.protocols.io/view/cambank-cfdna-bct-field-processing-v1-cyz2xx8e | Eliah G Overbey, Krista A Ryon, JangKeun Kim, Christopher E Mason | TITLE: CAMbank: cfDNA BCT Field Processing v1
AUTHORS: Eliah G Overbey, Krista A Ryon, JangKeun Kim, Christopher E Mason
[DESCRIPTION]
Field processing of cfDNA BCTs for the Cornell Aerospace Medicine Biobank (CAMbank).
Instructions for preserving: plasma and RBC Pellets.
[STEPS]
SECTION: Perform Venipuncture
1. Aft... | ["[Perform Venipuncture] After venipuncture, invert the tubes gently 8 to 10 times to fully mix tube anticoagulant with blood sample.\n\nStore the tube upright at room temperature until centrifugation.\n\nNote: The tube stabilizes cell-free DNA for up to 14 days at 6 °C to 37 °C and CTCs for up to 7 days at 15 °C to 30... |
62,535 | RMX Male Enhancement Reviews (Legit or Fake) Quality Ingredients That Work? | 3 | dx.doi.org/10.17504/protocols.io.n92ldz2qnv5b/v1 | https://www.protocols.io/view/rmx-male-enhancement-reviews-legit-or-fake-quality-b9bfr2jn | nonashinaty | TITLE: RMX Male Enhancement Reviews (Legit or Fake) Quality Ingredients That Work?
AUTHORS: nonashinaty
[DESCRIPTION]
RMX Male Enhancement Reviews
[STEPS] | [] |
89,748 | Yale Murine TMC - Tissue Preparation Protocol | 1 | dx.doi.org/10.17504/protocols.io.kqdg3x8keg25/v2 | https://www.protocols.io/view/yale-murine-tmc-tissue-preparation-protocol-c3vuyn6w | Yun-Hee Youm, Yufeng Liu, Vishwa Deep Dixit, Rong Fan, Bo Tao | TITLE: Yale Murine TMC - Tissue Preparation Protocol
AUTHORS: Yun-Hee Youm, Yufeng Liu, Vishwa Deep Dixit, Rong Fan, Bo Tao
[DESCRIPTION]
Protocol for embedding murine spleen, thymus and lymph node.
[STEPS]
SECTION: RNA Buffer (use before embedding tissue in OCT)
1. 1x PBS-MC Recipe
All prepared in DNAse RNAase EDTA... | ["[RNA Buffer (use before embedding tissue in OCT)] 1x PBS-MC Recipe\nAll prepared in DNAse RNAase EDTA free deionized water\nMgCl2 (1 mM, add 1 ml of 1 M MgCl2 stock solution into 1 L of 1x PBS)\nCaCl2 (0.1 mM, add 2 ml of 0.1 M of CaCl2 stock soution into 1 L of 1x PBS)\n4% paraformaldehyde (PFA), optional\n1 X Prote... |
null | null | null | dx.doi.org/10.17504/protocols.io.pbidike | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>This is a Single-cell RNA-Seq expression analysis <em>Support Protocol </em>for <a href="https://www.protocols.io/view/preparation-of-single-cell-rna-seq-libraries-for-n-n6gdhbw?step=27" target="_blank" rel="noopener noreferrer">Preparation of Single-Cell RNA-Seq Libraries fo... | [] |
81,804 | ABT263 (Navitoclax) Treatment in Mice | 4 | dx.doi.org/10.17504/protocols.io.5jyl8j4r6g2w/v1 | https://www.protocols.io/view/abt263-navitoclax-treatment-in-mice-ct5kwq4w | Andrew Davis, Peter Adams | TITLE: ABT263 (Navitoclax) Treatment in Mice
AUTHORS: Andrew Davis, Peter Adams
[DESCRIPTION]
This protocol is for treating mice with the senolytic, Navitoclax, by oral gavage.
[BEFORE_START]
ABT263 must first be dissolved in the vehicle by heating to 60°C. Sonication can be used to dissolve faster. See the material... | ["Give mice Navitoclax or Vehicle by oral gavage at 50mg/kg/day. Mice receive vehicle or drug for a total of 14 days, spread across 6 weeks, as outlined in the sub-steps below.", "Mice receive seven days of once-daily oral gavage of drug or vehicle", "Then fourteen days with no treatment", "Then seven more days of onc... |
27,990 | Chemically Competent Cells | null | dx.doi.org/10.17504/protocols.io.7jwhkpe | null | Alba Balletbó | TITLE: Chemically Competent Cells
AUTHORS: Alba Balletbó
[STEPS]
?. Grow to OD600 0.6 in 0.1/10 mL LB.
?. Centrifuge at 8000rpm for 5 minutes.
?. Resuspend in 5mL ice-cold CaCl2.
?. Distribute 1.5 mL over 3 Eppendorfs and spin for 30 seconds, 8000rpm.
?. Resuspend pellet in 0.5 mL ice-cold CaCl2, flick the tube gently... | ["Grow to OD600 0.6 in 0.1/10 mL LB.", "Centrifuge at 8000rpm for 5 minutes.", "Resuspend in 5mL ice-cold CaCl2.", "Distribute 1.5 mL over 3 Eppendorfs and spin for 30 seconds, 8000rpm.", "Resuspend pellet in 0.5 mL ice-cold CaCl2, flick the tube gently.", "Aliquot into 50µL Cryo-tubes and store at -80ºC until transfor... |
80,391 | Visualisation and quantification of dendritic spines in cultured human Medium Spiny Neurons (MSNs) | 1 | dx.doi.org/10.17504/protocols.io.261ge397jl47/v1 | https://www.protocols.io/view/visualisation-and-quantification-of-dendritic-spin-csrfwd3n | Quyen Do | TITLE: Visualisation and quantification of dendritic spines in cultured human Medium Spiny Neurons (MSNs)
AUTHORS: Quyen Do
[DESCRIPTION]
This protocol describes the visualisation of dendritic spines of human neurons cultured on coverslips in vitro and subsequent quantification using the software Imaris.
[STEPS]
SEC... | ["[Visualisation of fluorescent dendritic spines in MSNs] Mount a coverslip of Medium Spiny Neurons (MSNs) immunolabelled for DARP32 and neurobiotin onto SlowFade™ Diamond Antifade mountant (follow Protocol: Immunocytochemistry of cultured human Medium Spiny Neurons (MSNs)).", "[Visualisation of fluorescent dendritic s... |
49,018 | Preparation of DNA Samples for Copy Number Analysis by iDNA Technologies | 1 | null | https://www.protocols.io/view/preparation-of-dna-samples-for-copy-number-analysi-bt42nqye | Lynn Doran | TITLE: Preparation of DNA Samples for Copy Number Analysis by iDNA Technologies
AUTHORS: Lynn Doran
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Preparation of extracted genomic DNA for copy number analysis by </span><a href="https://www.idnagenetics.com/idna_location.php" style = "text-dec... | ["[Quote Preparation]\nEmail iDNA Technologies for a quote. Include the number of samples to be tested, which genes you want copy number analysis on, and which species the samples are from. Include one sample with a known copy number of 1 as an internal reference standard for each assay (gene).\nMinimum of 24 samples ... |
37,505 | Monoamine oxidase activity in fish brain tissue | 1 | dx.doi.org/10.17504/protocols.io.bgu9jwz6 | https://www.protocols.io/view/monoamine-oxidase-activity-in-fish-brain-tissue-bgu9jwz6 | Caio Maximino, Denis Broock Rosembeg | TITLE: Monoamine oxidase activity in fish brain tissue
AUTHORS: Caio Maximino, Denis Broock Rosembeg
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The protocol describes a spectrofluorophotometric method for rapid determination of monoamine oxidase (MAO) activity in zebrafish brains. The protocol... | ["[Reagent setup]\nPrepare tissue lysis buffer:16.8 mM Na2HPO4 and 10.6 mM KH2PO4, , isotonized with sucrose", "[Reagent setup]\nPrepare assay buffer:168 mM Na2HPO4 and 10.6 mM KH2PO4, , isotonized with KCl", "[Sample preparation]\nEuthanize fish in ice-cold water (Two brains should be pooled per to form one sample u... |
63,875 | PRIMARY NEURON CULTURE PROTOCOL | 4 | dx.doi.org/10.17504/protocols.io.14egn7jdmv5d/v1 | https://www.protocols.io/view/primary-neuron-culture-protocol-cambsc2n | Michael Lee | TITLE: PRIMARY NEURON CULTURE PROTOCOL
AUTHORS: Michael Lee
[DESCRIPTION]
This protocol details primary hippocampal neuron culture.
[BEFORE_START]
Before dissection
a. Dissection tools (scissors, forceps, spatulas, razor blades) should all be cleaned and autoclaved prior to use.
b. Prepare dishes or plates.
i. M... | ["[Primary Hippocampal Neuron Culture Protocol: Dissection] In laminar flow hood: have aluminum foil for mice, Kimwipes or paper towels for dissection, tools, ice bucket and Brain Bits Hibernate A (BB HA).", "[Primary Hippocampal Neuron Culture Protocol: Dissection] Begin dissection.", "[Primary Hippocampal Neuron Cult... |
33,964 | UPF study protocol | null | dx.doi.org/10.17504/protocols.io.bdeki3cw | https://www.protocols.io/view/upf-study-protocol-bdeki3cw | Ubaldo Bahemuka, Monica Kuteesa | TITLE: UPF study protocol
AUTHORS: Ubaldo Bahemuka, Monica Kuteesa
[STEPS] | [] |
52,732 | Image quantification | 4 | dx.doi.org/10.17504/protocols.io.bxq4pmyw | https://www.protocols.io/view/image-quantification-bxq4pmyw | Chunmei Chang | TITLE: Image quantification
AUTHORS: Chunmei Chang
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Image quantification</div></div>
[STEPS]
?. GUV image quantification
?. Analyze GUV images using a custom script implemented in Python 3.6 (https://github.com/Hurley-Lab/GUVquantification/blob/main/G... | ["GUV image quantification", "Analyze GUV images using a custom script implemented in Python 3.6 (https://github.com/Hurley-Lab/GUVquantification/blob/main/GUVintensity-2channel.ipynb).", "Bead image quantification", "Define the outline of individual bead based on the bright field channel.", "Calculate the intensity t... |
21,071 | UC Davis - Metabolomics: Sample preparation for Lipidomics | null | dx.doi.org/10.17504/protocols.io.ytpfwmn | null | Oliver Fiehn | TITLE: UC Davis - Metabolomics: Sample preparation for Lipidomics
AUTHORS: Oliver Fiehn
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block">This SOP describes sample extraction and sample preparation for lipid profiling by ... | ["Starting material: Plasma/serum: 30 µl sample volume or aliquot", "Sample Preparation: Switch on bath to pre-cool at –20°C (±2°C validity temperature range)Extraction solvents ♦ Purge both MeOH and MTBE for 5 min with nitrogen. ♦ Store solvents in the –20°C freezer to pre-chillHomogenization and extraction ♦... |
60,071 | Enhanced QIAseq DIRECT SARS-CoV-2 Kit for Illumina MiSeq | 4 | dx.doi.org/10.17504/protocols.io.rm7vzy39rlx1/v3 | https://www.protocols.io/view/enhanced-qiaseq-direct-sars-cov-2-kit-for-illumina-b6wfrfbn | Padmini Ramachandran, Tamara Walsky, Amanda Windsor, Maria Hoffmann, Chris Grim | TITLE: Enhanced QIAseq DIRECT SARS-CoV-2 Kit for Illumina MiSeq
AUTHORS: Padmini Ramachandran, Tamara Walsky, Amanda Windsor, Maria Hoffmann, Chris Grim
[DESCRIPTION]
This SARS-CoV-2 targeted, a rapid viral RNA library preparation kit compatible with Illumina sequencers, provides an approximately five hour tur... | ["[cDNA Synthesis Procedure]", "[cDNA Synthesis Procedure] Thaw template RNA on ice. Gently yet thoroughly mix, then briefly centrifuge to collect residual liquid from the sides of the tubes and return to ice.", "[cDNA Synthesis Procedure] While keeping reagents on ice, prepare the cDNA reaction according to the table ... |
43,630 | Shipping beetles to the UF Forest Entomology Lab | 3 | dx.doi.org/10.17504/protocols.io.bnunmeve | https://www.protocols.io/view/shipping-beetles-to-the-uf-forest-entomology-lab-bnunmeve | Jiri Hulcr | TITLE: Shipping beetles to the UF Forest Entomology Lab
AUTHORS: Jiri Hulcr
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes how to send specimens to the UF Forest Entomology Lab.</div><div class = "text-block"><span>This protocol is part of the Bark Beetle Mycobiome (BBM) Res... | [] |
91,700 | TF and epigenetic modifier CRISPRi/a screens in human T cells | 4 | null | https://www.protocols.io/view/tf-and-epigenetic-modifier-crispri-a-screens-in-hu-c5suy6ew | Andrea R Daniel | TITLE: TF and epigenetic modifier CRISPRi/a screens in human T cells
AUTHORS: Andrea R Daniel
[DESCRIPTION]
This protocol describes methods for CRISPR interference or activation screens identifying transcriptional and epigenetic regulators of human CD8+ T cell state.
[STEPS]
SECTION: TF and epi-modifier CRISPRi/a gR... | ["[TF and epi-modifier CRISPRi/a gRNA library construction] The TSSs for each TF and epi-modifier were extracted using CRISPick and 1,000-bp windows were constructed around each TSS (−500 to +500 bp).", "[TF and epi-modifier CRISPRi/a gRNA library construction] After establishing an SaCas9 gRNA database with the strict... |
71,160 | Anthropometry (Standing Height) | 1 | dx.doi.org/10.17504/protocols.io.yxmvm2kzog3p/v1 | https://www.protocols.click/view/anthropometry-standing-height-chqyt5xw | g.spencer | TITLE: Anthropometry (Standing Height)
AUTHORS: g.spencer
[DESCRIPTION]
Standing height is an assessment of maximum vertical size. Take this measure on all patients aged 6 years and older who are able to stand unassisted. Standing height is measured with a fixed stadiometer with a vertical backboard and a moveable hea... | ["[Equipment Required]", "[Preparing for Measurement] Have the patient remove their footwear, hair accessories or hats from the top of their head", "[Standing Height Measurement] 1. Ask the patient to stand erect, arms hanging at their sides, feet together, heels, buttocks and back in contact with the stadiometer.\n\n2... |
50,899 | Protocol for Mouse IP Anesthesia (Ketamine-Xylazine) | 4 | dx.doi.org/10.17504/protocols.io.bvxtn7nn | https://www.protocols.io/view/protocol-for-mouse-ip-anesthesia-ketamine-xylazine-bvxtn7nn | Haowei Li, Markus Holzl, Mohsen Khosravi-Maharlooei, Nichole Danzl, Austin Chen, Megan Sykes | TITLE: Protocol for Mouse IP Anesthesia (Ketamine-Xylazine)
AUTHORS: Haowei Li, Markus Holzl, Mohsen Khosravi-Maharlooei, Nichole Danzl, Austin Chen, Megan Sykes
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol details the constitution of our anesthesia cocktail that we use for sedating... | ["[For Intraperitoneal (IP) Approach:]\nUse 10 uL/gram of anesthesiaComments:1. It takes time until the mice fall asleep 2. Very often, mice do not fall asleep but move around like zombies. This requires another small dose of IP ketamine-xylazine injection or a small dose of IV injectiona. When doing IP injection again... |
95,767 | GENERATION OF LYSATE INOCULATION MATERIALS | 1 | dx.doi.org/10.17504/protocols.io.4r3l22d2ql1y/v1 | https://www.protocols.io/view/generation-of-lysate-inoculation-materials-c9rxz57n | Scott Vermilyea | TITLE: GENERATION OF LYSATE INOCULATION MATERIALS
AUTHORS: Scott Vermilyea
[DESCRIPTION]
This protocol details inoculation lysate preparation.
[STEPS]
SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
1. Acquire the tissue from 4-month-old asymptomatic TgA53T (Line G2-3) and e... | ["[Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue] Acquire the tissue from 4-month-old asymptomatic TgA53T (Line G2-3) and end-stage tissue harvest and store at -80 °C prior to use.", "[Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue] Weigh the tissue ... |
67,383 | Diaetoxil | 1 | dx.doi.org/10.17504/protocols.io.n92ldzdonv5b/v1 | https://www.protocols.io/view/diaetoxil-cd2xs8fn | natedif | TITLE: Diaetoxil
AUTHORS: natedif
[DESCRIPTION]
➢ Produktname – Diaetoxil
➢ Nebenwirkungen – NA
➢ Zusammensetzung — Natürliche organische Verbindung
➢ Vorteile – Hilft, Gewicht zu reduzieren und die Energie zu steigern!
➢ Verfügbarkeit — Auf Lager
➢ Bewertung —⭐⭐⭐⭐⭐
➢ Preis – Klicken Sie hier
➢ (Verkauf läuft) – htt... | [] |
104,801 | Obtaining Competent Cells | 0 | dx.doi.org/10.17504/protocols.io.j8nlk86z1l5r/v1 | https://www.protocols.io/view/obtaining-competent-cells-dij94cr6 | Miquel Estévez-Gay | TITLE: Obtaining Competent Cells
AUTHORS: Miquel Estévez-Gay
[DESCRIPTION]
Obtencion of 100ml competent E.coli aliquots that must be stored in -80C
[GUIDELINES]
Competent cells are fragile. All work must be done under a laminar flux cabin and in aseptic conditions.
[STEPS]
1. Incubate 10ml of E.coli BL21(DE3) or DH... | ["Incubate 10ml of E.coli BL21(DE3) or DH5α o/n without plasmid (from –80°C stock). To do that, add 10 mL of sterile LB media in a 50ml sterile falcon tube. With the use of a yellow micropipette, scratch the surface of the frozen E.coli from the -80°C freezer and toss it into the Falcon containing the media. This must ... |
20,482 | Thawing Rosettes and NPC | null | dx.doi.org/10.17504/protocols.io.x9afr2e | null | Celeste Karch, Rita Martinez, Jacob Marsh | TITLE: Thawing Rosettes and NPC
AUTHORS: Celeste Karch, Rita Martinez, Jacob Marsh
[STEPS]
?. Aliquot of pre-warmed DMEM/F12 into a 15 ml conical tube.
9 ml
?. Remove cells from liquid nitrogen and thaw in water bath for approximately or until a small ice clump remains.
?. Add freshly thawed cells into theof pre-war... | ["Aliquot of pre-warmed DMEM/F12 into a 15 ml conical tube.\n9 ml", "Remove cells from liquid nitrogen and thaw in water bath for approximately or until a small ice clump remains.", "Add freshly thawed cells into theof pre-warmed DMEM/F12 and mix gently by gently shaking tube 3 times. Avoid breaking up clumps of cell... |
null | null | null | dx.doi.org/10.17504/protocols.io.fv4bn8w | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p><em>The EpiQuik™ Total Histone Extraction Kit</em> is a complete set of optimized buffers and reagents for extracting total core histone proteins (H2A, H2B, H3, and H4) from mammalian cells or tissues in a simple 60 minute procedure. The post-translational modifications (PTM)... | [] |
null | null | null | dx.doi.org/10.17504/protocols.io.e7bbhin | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>Using UV-induced peptide exchange, MHC/peptide monomers can be generated with conditional Flex-T™ monomers that harbor peptides of interest in their binding grooves. These new MHC monomers are subsequently multimerized using streptavidin-fluorophore conjugates. The resulting ... | [] |
99,427 | WATER PRODUCTION FOR AWARE (Organic Contaminants) | 0 | dx.doi.org/10.17504/protocols.io.14egn67jql5d/v1 | https://www.protocols.io/view/water-production-for-aware-organic-contaminants-ddcb22sn | Celia Manaia | TITLE: WATER PRODUCTION FOR AWARE (Organic Contaminants)
AUTHORS: Celia Manaia
[DESCRIPTION]
The protocol summarises the procedures used for analytical control. The protocol describes the Standard Operating Procedure (SOP) for the optimization of advanced tertiary treatment of water, based on a comprehensive quality a... | ["[Organic Contaminants:] The water production for AWARE main activities includes three stages – disinfection by ultraviolet C radiation (UVC), storage for720 min-1440 min(according to water load and season) and ozonation. The water quality is monitored at these three stages, for the parameters indicated in Figure 1 be... |
69,321 | Green Lab Nanoparticle For 6 Well Cardiomyocyte Transfection | 4 | dx.doi.org/10.17504/protocols.io.ewov1ojd2lr2/v1 | https://www.protocols.io/view/green-lab-nanoparticle-for-6-well-cardiomyocyte-tr-cfxhtpj6 | Edwin Yoo | TITLE: Green Lab Nanoparticle For 6 Well Cardiomyocyte Transfection
AUTHORS: Edwin Yoo
[DESCRIPTION]
nanoparticle transfection
[STEPS]
SECTION: Nanoparticle Synthesis
1. DNA Dilution
For 3 replicate wells of one condition in 6-well plate format, mix the following in microcentrifuge tube.
60 ul DNA at 1ug/ul
4... | ["[Nanoparticle Synthesis] DNA Dilution\nFor 3 replicate wells of one condition in 6-well plate format, mix the following in microcentrifuge tube.\n60 ul DNA at 1ug/ul\n440 ul sodium acetate\nTotal volume = 500 ul for 3 replicate wells per one condition", "[Nanoparticle Synthesis] Polymer Dilution (7.2 mg.ml polymer co... |
null | null | null | dx.doi.org/10.17504/protocols.io.ftqbnmw | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>Defined Medium for Aureococcus anophagefferens</p>
[GUIDELINES]
<table>
<tbody>
<tr style="text-align: center;">
<td colspan="7">
<p><strong>Anhydrous Salts</strong></p>
</td>
</tr>
<tr style="text-align: center;">
<td>
<p><strong>Constituent</strong></p>
</td>
<td>
<p><stro... | [] |
19,994 | 0.5 M EDTA (0.5 L) | null | dx.doi.org/10.17504/protocols.io.xr2fm8e | null | Mattia Pegoraro | TITLE: 0.5 M EDTA (0.5 L)
AUTHORS: Mattia Pegoraro
[DESCRIPTION]
<p>Is used in a multitude of experiments, but is often used as an ingredient in 10x TBE buffer</p>
[STEPS]
?. Fill 0.5 L beker with of deionized water.
300 ml
?. Add of Ethylenediamine Tetraacetic Acid Disodium Salt (292,24 g/mol) to the water and mix.
... | ["Fill 0.5 L beker with of deionized water.\n300 ml", "Add of Ethylenediamine Tetraacetic Acid Disodium Salt (292,24 g/mol) to the water and mix.\n73 g", "Add NaOH pellets until the pH is near 7.8 (the solution will be clear around pH 7.5). Complete to arrive at pH 8 with NaOH 5 M solution (also NaOH 10 M is good).", "... |
null | null | null | dx.doi.org/10.17504/protocols.io.r5ud86w | null | null | TITLE: No Title
AUTHORS:
[GUIDELINES]
<p>Keeps cells always on ice & all reagents must be ice-cold</p>
<p> </p>
[STEPS]
?.
?.
?.
?.
?.
?.
?.
?.
?.
?.
?.
?. | [] |
52,126 | Glucagon measurement from islet populations using a TR-FRET assay | 1 | dx.doi.org/10.17504/protocols.io.rm7vz3r74gx1/v1 | https://www.protocols.io/view/glucagon-measurement-from-islet-populations-using-bw56pg9e | Wesley Eaton, Michael Roper | TITLE: Glucagon measurement from islet populations using a TR-FRET assay
AUTHORS: Wesley Eaton, Michael Roper
[DESCRIPTION]
TR-FRET homogenous assay method using a microfluidic device for quantitation of glucagon secretion from human islet populations is described. A PDMS microfluidic device was used to perfuse a po... | ["Aquire Chemicals and Reagents", "[Chemical reagents] TR-FRET glucagon assay -Cisbio (Walthan, MA)", "Polydimethylsiloxane (PDMS) prepolymer (Sylgard 184) -Dow Corning (Midland, MI)", "[Chemical reagents] Dextrose -Fisher Scientific (Pittsburg, PA)", "[Chemical reagents] Polydimethylsiloxane (PDMS) prepolymer (Sylgard... |
94,531 | Electrophysiology with iPSC-derived neurons | 4 | dx.doi.org/10.17504/protocols.io.q26g7pebkgwz/v1 | https://www.protocols.io/view/electrophysiology-with-ipsc-derived-neurons-c8jbzuin | Minee-Liane Choi | TITLE: Electrophysiology with iPSC-derived neurons
AUTHORS: Minee-Liane Choi
[DESCRIPTION]
This protocol describes the method to perform patch-clamp recordings of iPSC-derived neurons.
[STEPS]
1. Patch-clamp recordings of iPSC-derived neurons are performed using an infrared differential interference contrast (DIC) i... | ["Patch-clamp recordings of iPSC-derived neurons are performed using an infrared differential interference contrast (DIC) imaging system on an Olympus BX51WI upright microscope (Olympus, Japan) coupled with a Multipatch 700B amplifier under the control of pClamp 10.2 software package (Molecular Devices, USA)", "For the... |
41,635 | VCF2PCP | 5 | dx.doi.org/10.17504/protocols.io.bkwbkxan | https://www.protocols.io/view/vcf2pcp-bkwbkxan | Judith Ballesteros Villascan, Israel Aguilar Ordoñez, Fernando Pérez-Villatoro | TITLE: VCF2PCP
AUTHORS: Judith Ballesteros Villascan, Israel Aguilar Ordoñez, Fernando Pérez-Villatoro
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Nextflow pipeline that runs and plots admixture and smartpca from a compressed VCF.</div></div>
[STEPS]
?. [Pre-processing]
Split chromosomesSplit c... | ["[Pre-processing]\nSplit chromosomesSplit chromosomes from a compressed VCF file.Dependencies:", "[Pre-processing]\nSimplify and remove LDSimplifyVCCF to keep only INFO/AF and GT and removes LD variants with bcftools +prune. Please, consider window for LD pruning is given in bp.Dependencies:\na) Remove variants in LD.... |
36,633 | Slice Culture Media | null | dx.doi.org/10.17504/protocols.io.bfzzjp76 | https://www.protocols.io/view/slice-culture-media-bfzzjp76 | Allen Institute for Brain Science | TITLE: Slice Culture Media
AUTHORS: Allen Institute for Brain Science
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes the preparation of Slice Prep Media used for human or mouse slice culture.</div><div class = "text-block"><span style = "font-weight:bold;">Note</span><span>:... | [] |
104,396 | Chronic Multi-Electrode Assembly Implantation with a Chamber Setup | 0 | dx.doi.org/10.17504/protocols.io.bp2l62m95gqe/v1 | https://www.protocols.io/view/chronic-multi-electrode-assembly-implantation-with-dh7k39kw | Jiwon Choi, Usamma Amjad, Helen N Schwerdt | TITLE: Chronic Multi-Electrode Assembly Implantation with a Chamber Setup
AUTHORS: Jiwon Choi, Usamma Amjad, Helen N Schwerdt
[DESCRIPTION]
This protocol describes how to implant an array of carbon fiber and other types of microelectrodes for chronic recording using the two-part chamber system. The previous steps to i... | ["[Setup for Multi-Electrode Assembly Installation] Procedures were performed on Rhesus monkeys (n = 2) and were approved by the Institute’s Animal Care and Use Committee (IACUC) at the University of Pittsburgh and were performed following the Guide for the Care and Use of Laboratory Animals (Department of Health and H... |
18,040 | A Breif Look of Phage Display Technology | null | dx.doi.org/10.17504/protocols.io.vuye6xw | null | bello smitu | TITLE: A Breif Look of Phage Display Technology
AUTHORS: bello smitu
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">A Breif Look of Phage Display Technology</div></div><div class = "text-block"><div class = "justify" style = "text-align:left"><span s... | [] |
89,960 | Cryo-EM sample preparation for full length LRRK2(I2020T):MLi-2/GZD-824-E11 DARPin complex | 1 | dx.doi.org/10.17504/protocols.io.yxmvm35n9l3p/v1 | https://www.protocols.io/view/cryo-em-sample-preparation-for-full-length-lrrk2-i-c34gyqtw | Marta Sanz Murillo, Amalia Villagran Suarez | TITLE: Cryo-EM sample preparation for full length LRRK2(I2020T):MLi-2/GZD-824-E11 DARPin complex
AUTHORS: Marta Sanz Murillo, Amalia Villagran Suarez
[DESCRIPTION]
Guide for cryo-EM grid preparation of FL-LRRK2 bound to Type-I/Type-II inhibitors
[BEFORE_START]
Before starting cryo-EM grid preparation section: Set up ... | ["[Protein purification] His6-Z-TEV-LRRK2 was expressed and purified as described in a previous protocol", "[Sample preparation] Prepare LRRK2 buffer. Keep it at 4ºC.\n20 millimolar (mM) HEPES pH=7.4\n150 millimolar (mM) NaCl\n2.5 millimolar (mM) MgCl2\n20 micromolar (µM) GDP\n0.5 millimolar (mM) TCEP", "[Sample prepar... |
99,446 | Apomorphine-induced rotations | 0 | dx.doi.org/10.17504/protocols.io.x54v92yozl3e/v1 | https://www.protocols.io/view/apomorphine-induced-rotations-ddcw22xe | Santiago Unda, Mihaela Stavarache, Michael G. Kaplitt, Roberta Marongiu | TITLE: Apomorphine-induced rotations
AUTHORS: Santiago Unda, Mihaela Stavarache, Michael G. Kaplitt, Roberta Marongiu
[DESCRIPTION]
To test for 6-OHDA lesion efficiency, rotational behaviors are assessed while mice are under the effects of apomorphine. This test is run during the dark phase of light cycle. Experiments... | ["Prepare apomorphine in 0.9% saline/0.1% ascorbate solution to dose each mouse with 0.25mg/kg via subcutaneous injection.\n*Note that this solution is light-sensitive and should be shielded from light.", "[Preparation] Weigh mice and habituate to the testing room for at least 1 hour prior to the start of testing.", "C... |
null | null | null | dx.doi.org/10.17504/protocols.io.iqycdxw | null | null | TITLE: No Title
AUTHORS:
[DESCRIPTION]
<p>Procedure for DNA extractions using PowerWater DNA extraction kit from filtered LEO seepage water samples and prelimnary QC by nanodrop</p>
[BEFORE_START]
<p><strong>A) Identify samples to extract</strong></p>
<p>Determine <span style="text-decoration: underline;">number of ... | [] |
20,321 | Tranfection of sgRNA using SpCas9 containing plasmids to generate cell lines with Cas9 | null | dx.doi.org/10.17504/protocols.io.x39fqr6 | null | Binnypreet Kaur, Drahomíra Faktorová, Julius Lukeš, , | TITLE: Tranfection of sgRNA using SpCas9 containing plasmids to generate cell lines with Cas9
AUTHORS: Binnypreet Kaur, Drahomíra Faktorová, Julius Lukeš, ,
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">This protocol describes transfection of r... | ["Step1: Assemble RNP complexes with (30pmol/μl) of gRNA and (20pmol/μl) of Cas9 in the ratio of (Cas9:gRNA) 1:9 in resuspension buffer before electroporation and keep it at room temperature for 10 min and then transfer it to ice untill use.", "Step2: Follow the protocol 7 for transfection"] |
42,914 | Molecular Cloning | 1 | dx.doi.org/10.17504/protocols.io.e6nvw59o9vmk/v1 | https://www.protocols.io/view/molecular-cloning-bm6ak9ae | Lizhen Zhu, Ruixin Lin, Xiaoqi Ye, Ruiyan zeng, 18814306949 Minkun Huang, 624787660 | TITLE: Molecular Cloning
AUTHORS: Lizhen Zhu, Ruixin Lin, Xiaoqi Ye, Ruiyan zeng, 18814306949 Minkun Huang, 624787660
[DESCRIPTION]
SZPT-CHINA Experimental methods of molecular cloning
[GUIDELINES]
advisers WeiJia Wu
LiJun Zhang（PhD)
Yongjun Tang(PhD)
[STEPS]
SECTION: Vector linearization and target fragment prepara... | ["[Vector linearization and target fragment preparation] Q5 DNA polymerase PCR system(50 µL )\n 25 µL \n2 µL \n2 µL \n1 µL \n20 µL \n \nPCR Program", "[Enzyme digestion Connection] Enzyme digestion Connection", "[Transformation] Transformation", "[Transformation] Take 50 µL of competent cells in a sterile Eppendorf tub... |
30,303 | Isolation of supernumerary mini-chromosomes from fungi for enrichment sequencing | null | dx.doi.org/10.17504/protocols.io.9t7h6rn | null | Thorsten Langner, Adeline Harant, Sophien Kamoun | TITLE: Isolation of supernumerary mini-chromosomes from fungi for enrichment sequencing
AUTHORS: Thorsten Langner, Adeline Harant, Sophien Kamoun
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Fungal genomes are highly dynamic and often contain supernumerary mini-chromosomes. However, our knowledge... | ["[Preparation of the protoplasts]\nPrepare a fresh culture of M. oryzae on complete medium agar plates.", "[Preparation of the protoplasts]\nIncubate at 24°C for 7 days.", "[Preparation of the protoplasts]\nInoculate 150 ml of YG medium in a 500 ml Erlenmeyer flask with 8 pieces (~0.5 x 0.5 cm) of 7 days old M. oryzae... |
52,863 | Concentration of viruses from sewage using HA filters | 4 | dx.doi.org/10.17504/protocols.io.bxu7pnzn | https://www.protocols.io/view/concentration-of-viruses-from-sewage-using-ha-filt-bxu7pnzn | Adélaïde Roguet, Shuchen Feng | TITLE: Concentration of viruses from sewage using HA filters
AUTHORS: Adélaïde Roguet, Shuchen Feng
[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Concentration of viruses from sewage using HA filters</span><span style = "font-weight:bold;"></span></div></div>
[ST... | ["[Filtration of the sewage samples (in the biosafety cabinet)]\nTake the sample out from the refrigerator.", "[Filtration of the sewage samples (in the biosafety cabinet)]\nHomogenize the sample thoroughly, avoiding foaming.", "[Filtration of the sewage samples (in the biosafety cabinet)]\nUse a pipette controller, tr... |
94,181 | LC/MS analysis of plasma samples from PPMI | 4 | dx.doi.org/10.17504/protocols.io.81wgbxe2ylpk/v1 | https://www.protocols.io/view/lc-ms-analysis-of-plasma-samples-from-ppmi-c78dzrs6 | John Chen, Romeo Maciuca, Paul Benton, Jung Suh, Sonnet Davis, Sarah Huntwork-Rodriguez, Ning Xia, Michael A. Schwarzschild | TITLE: LC/MS analysis of plasma samples from PPMI
AUTHORS: John Chen, Romeo Maciuca, Paul Benton, Jung Suh, Sonnet Davis, Sarah Huntwork-Rodriguez, Ning Xia, Michael A. Schwarzschild
[DESCRIPTION]
Plasma samples from PPMI were analyzed by liquid chromatography with mass spectrometry (LC/MS) for a variety of metabolite... | ["[Plasma sample preparation for LC/MS analysis] Thawed plasma samples were spun down at 3.5k*g for 10 min at 4°C.", "[Plasma sample preparation for LC/MS analysis] Plasma (10 µL) samples were transferred to an Agilent Captiva collection plate and processed via the Agilent Bravo Metabolomics Sample Prep Platform (Agile... |
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