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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
Increased sensitivity of Euplotes crassus to selective agents using 0.3 M glucose-based culture conditions.
 
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Lawrence A. Klobutcher 10.17504/protocols.io.gwnbxde Protist Research to Optimize Tools in Genetics (PROT-G) Numerous unpublished studies indicate that Euplotes crassus is resistant to extremely high concentrations of commonly used selective agents when grow in artificial seawater. We have found that when E. crassus is grown in 1 part artificial seawater + 9 parts 0.3 M glucose, they are much more sensitive to a number of selective agents. For example, 1080 ug/ml paromomycin in artificial seawater only inhibited growth, as little as 120 ug/ml was effective in killing cells when grown with 0.3 M glucose. Similarly, G418 at 400 ug/ml had little effect in seawater, but as little as 100 ug/ml blocked growth in 0.3 M glucose. This may not prove true for all selective agents, as we saw no effect with paclitaxel up to 100 uM under either growth condition. University of Connecticut Health Center 1 2017 Lawrence A. Klobutcher 2017. Increased sensitivity of Euplotes crassus to selective agents using 0.3 M glucose-based culture conditions.. protocols.io dx.doi.org/10.17504/protocols.io.gwnbxde 2021-03-29 03:09:51
ChroPlate - ProteinG
 
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Alexandra Ehl, David Frommholz, Nadine Stefanczyk 10.17504/protocols.io.uyjexun Purification Guide for the Isolation of Antibodies with ChroPlate Filtration Plates by DALEX Biotech.Easy and quick high throughput antibody purification from various sources and species.Each well of the ChroPlate has a binding capacity of > 1 mg (tested with human polyclonal Ig, binding varies between species and clones).The proprietary resin does not shrink or swell in aqueous buffers.High pressure stability.pH stability short term 2 - 8, long term 3 - 8.Excellent thermal stability up to 15 minutes at 80 °C in aqueous buffers at neutral pH.Can be dried for long term storage (80 °C for > 2 h). DALEX Biotech, DALEX Biotech, DALEX Biotech https://dalex-biotech.com/ 1 2018 Alexandra Ehl, David Frommholz, Nadine Stefanczyk 2018. ChroPlate - ProteinG. protocols.io dx.doi.org/10.17504/protocols.io.uyjexun 2021-03-29 03:09:53
Preparation of PCR amplicons from edited cells for deep sequencing
 
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Mark DeWitt 10.17504/protocols.io.hdsb26e CornLab This protocol describes how to extract and amplify DNA from genome-edited cells, and sequence the resulting amplicons using Illumina MiSeq NGS. Innovative Genomics Institute 1 2017 Mark DeWitt 2017. Preparation of PCR amplicons from edited cells for deep sequencing. protocols.io dx.doi.org/10.17504/protocols.io.hdsb26e 2021-03-29 03:09:52
Colony PCR
 
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宏亮 董 10.17504/protocols.io.7h2hj8e 2019 iGEM NEFU_China This PCR method can be used to screen for inserted target genes or DNA sequencing analysis. Northeast Forest University 1 2019 宏亮 董 2019. Colony PCR. protocols.io dx.doi.org/10.17504/protocols.io.7h2hj8e 2021-03-29 03:09:51
High-Throughput Beta-glucuronidase (GUS) assay for Phaeodactylum tricornutum
 
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Erin Garza, Vincent Bielinski 10.17504/protocols.io.bbexijfn Protist Research to Optimize Tools in Genetics (PROT-G), JCVI West Protocols A high-throughput method for measuring β-glucuronidase (GUS) activity in the diatom Phaeodactylum tricornutum. This protocol has been optimized for 250 μl volumes. For larger volumes see the following protocol dx.doi.org/10.17504/protocols.io.hefb3bn, which this protocol was based off of. J. Craig Venter Institute, J. Craig Venter Institute, Synthetic Biology & Bioenergy Group 1 2020 Erin Garza, Vincent Bielinski 2020. High-Throughput Beta-glucuronidase (GUS) assay for Phaeodactylum tricornutum. protocols.io dx.doi.org/10.17504/protocols.io.bbexijfn 2021-03-29 03:09:50
Viability Estimation of Islets for Distribution Using Inclusion and Exclusion Fluorescent Dyes (FDA/PI)
 
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Integrated Islet Distribution Program 10.17504/protocols.io.43kgykw Integrated Islet Distribution Program This Standard Operating Procedure is adapted from the work of the 'National Institutes of Health-Sponsored Clinical Islet Transplantation Consortium Phase 3 Trial: Manufacture of a Complex Cellular Product at Eight Processing Facilities'' following the SOP cited in the document 'Purified Human Pancreatic Islet - Viability Estimation of Islet Using Fluorescent Dyes (FDA/PI): Standard Operating Procedure of the NIH Clinical Islet Transplantation Consortium'This SOP defines the procedure for assessment of viability of human isolated islet preparations, which include endocrine and exocrine tissue, for use in the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) sponsored research in the Integrated Islet Distribution Program (IIDP). This protocol is written to assist the participating islet isolation centers and investigators who are part of this program.Fluorescein Diacetate/ Propidium Iodide (FDA)/(PI) Viability Assay is a rapid fluorometric method to test the integrity of the plasma membrane simultaneously using inclusion and exclusion dyes; the assay differentiates between viable and nonviable cells and is, consequently, used for determination of viability of islet preparations. Integrated Islet Distribution Program, City of Hope, Duarte, CA 1 2019 Integrated Islet Distribution Program 2019. Viability Estimation of Islets for Distribution Using Inclusion and Exclusion Fluorescent Dyes (FDA/PI). protocols.io dx.doi.org/10.17504/protocols.io.43kgykw 2021-03-29 03:09:55
Pinpoint Science 30-Second Covid-19 Test Protocol
 
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lisa.diamond 10.17504/protocols.io.bkxmkxk6 XPRIZE Rapid Covid Testing Pinpoint's nanosensor-based platform detects SARS-CoV-2 antigen by (1) measuring baseline current through our functionalized nanosensor chip in ordinary PBS buffer; (2) measuring the change in current at specific voltage ranges after insertion of the specimen as a result of binding between probe aptamer and target antigen; (3) computing delta between these two measurements. These measurements and computations are performed internally in quadruplicate, with four separately measured nanosensor channels. Consolidated results are reported to the user as {Detected; Not Detected; Inconclusive; or Error}. Pinpoint Science Inc. 1 2020 lisa.diamond 2020. Pinpoint Science 30-Second Covid-19 Test Protocol. protocols.io dx.doi.org/10.17504/protocols.io.bkxmkxk6 2021-03-29 03:09:51
UC Davis - Metabolomics: Primary metabolites by GC-TOF MS
 
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Oliver Fiehn 10.17504/protocols.io.64nhgve Mouse Metabolic Phenotyping Centers, Metabolomics Protocols & Workflows Description of standard setting for analysis of derivatized samples ready for injection into a Leco Pegasus III or Pegasus IV gas chromatograph-time of flight mass spectrometer.References: Fiehn O, Wohlgemuth G, Scholz M, Kind T, Lee DY, Lu Y, Moon S, Nikolau BJ (2008) Quality control for plant metabolomics: Reporting MSI-compliant studies. Plant J. 53, 691-704 University of California, Davis https://mmpc.org/shared/document.aspx?id=83&docType=Protocol 2 2019 Oliver Fiehn 2019. UC Davis - Metabolomics: Primary metabolites by GC-TOF MS. protocols.io dx.doi.org/10.17504/protocols.io.64nhgve 2021-03-29 03:09:51
Aegea Biotechnologies rapid PCR SARS-CoV-2 test (high sensitivity & specificity; able to detect different strain types)
 
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Lyle J. Arnold Ph.D., Stella M. Sung Ph.D. 10.17504/protocols.io.bk2dkya6 Coronavirus Method Development Community, XPRIZE Rapid Covid Testing This protocol is for the Aegea Biotechnologies rapid PCR-based SARS-CoV-2 test. This assay uses patented "Switch-Blocker" technology as well as taqman probes to test for presence of SARS-CoV-2 and simultaneously orthogonally validate. A single amplification reaction is performed, and "Switch Blocker" is used on the forward strand and taqman is used on the reverse strand. The assay design has high sensitivity & specificity--single nucleotide level. Moreover, it is able to detect the SARS-CoV-2 L strain vs the SARS-CoV-2 S strain. The test can be adapted to point of care (Roche LIAT) as well as for different SARS-CoV-2 strains as the virus mutates. A next generation version of the assay could identify the presence of the SARS-CoV-2 L/S strains vs. influenza A/B. Finally, because of the sensitivity and specificity, the Aegea PCR-based SARS-CoV-2 test should be able to use saliva samples, and it is suitable for pooled testing. This protocol is designed for high throughput PCR (96 or 384 well plate formats).Keywords: PCR, COVID-19, coronavirus, SARS-CoV-2, high throughput, multiplex, Switch-Blocker, taqman, high sensitivity, high specificity, accurate, pooling, saliva, strain types, L-strain, S-strain, combination SARS-CoV-2 and influenza Aegea Biotechnologies, Inc. http://www.aegeabiotech.com 1 2020 Lyle J. Arnold Ph.D., Stella M. Sung Ph.D. 2020. Aegea Biotechnologies rapid PCR SARS-CoV-2 test (high sensitivity & specificity; able to detect different strain types). protocols.io dx.doi.org/10.17504/protocols.io.bk2dkya6 2021-03-29 03:09:52
Fruit fly potato culture media
 
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Jorge Frias, Nelson Simões 10.17504/protocols.io.bei3jcgn Potato-based medium for culture of Drosophila melanogaster. CBA – Centre for Biotechnology of Azores, University of Azores, 9500- 321 Ponta Delgada, Açores. Portugal., CBA – Centre for Biotechnology of Azores, University of Azores, 9500- 321 Ponta Delgada, Açores. Portugal. 1 2020 Jorge Frias, Nelson Simões 2020. Fruit fly potato culture media. protocols.io dx.doi.org/10.17504/protocols.io.bei3jcgn 2021-03-29 03:09:52
DNA extraction from sputum
 
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Laszlo Irinyi, Lana Pasic, Wieland Meyer 10.17504/protocols.io.6enhbde Westmead Institute for Medical Research, The University of Sydney, The University of Sydney 1 2019 Laszlo Irinyi, Lana Pasic, Wieland Meyer 2019. DNA extraction from sputum. protocols.io dx.doi.org/10.17504/protocols.io.6enhbde 2021-03-29 03:09:52
U Michigan - Sirius Red staining
 
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Jeff Hodgin 10.17504/protocols.io.56tg9en Mouse Metabolic Phenotyping Centers Picrosirius red staining (also called Sirius red staining) highlights fibrosis by staining collagen (collagen 1 and III fibers) fibers in paraffin-embedded kidney tissue sections. Sirius red stained collagen appears red by light microscopy. When polarized, the birefringence is highly specific for collagen. The larger collagen fibers are bright yellow or orange, and the thinner ones, including reticular fibers, are green. University of Michigan - Ann Arbor https://mmpc.org/shared/document.aspx?id=313&docType=Protocol 2 2019 Jeff Hodgin 2019. U Michigan - Sirius Red staining. protocols.io dx.doi.org/10.17504/protocols.io.56tg9en 2021-03-29 03:09:55
Plasmid Modification by Annealed Oligo Cloning
 
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Addgene The Nonprofit Plasmid Repository 10.17504/protocols.io.bawkifcw This protocol describes plasmid modification by annealed oligo cloning. To see the full abstract and additional resources, please visit the Addgene protocol page. Addgene https://www.addgene.org/protocols/annealed-oligo-cloning/ 2 2019 Addgene The Nonprofit Plasmid Repository 2019. Plasmid Modification by Annealed Oligo Cloning. protocols.io dx.doi.org/10.17504/protocols.io.bawkifcw 2021-03-29 03:09:52
HBV DNA Real Time Quantification according to ARNS 12187 project
 
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Berthold Bivigou-Mboumba, Sandrine François-Souquière, Luc Deleplancque, Jeanne Sica, Augustin Mouinga-Ondémé, Marie Amougou-Atsama, Marie Laure Chaix, Richard Njouom, François Rouet 10.17504/protocols.io.kw4cxgw HBV DNA Real Time Quantification according to ARNS 12187 projectUsing the Arrow extractor (NorDiag, Biotrin International, Ireland), DNA was extracted from 240 μl of plasma, pre-treated with10 μl of proteinase K, using the Arrow Viral NA extraction kit, according to the manufacturer’s instructions during 45 min. Template DNA was eluted into 60 μl of kit elution buffer. Besides clinical samples, quantification standard (Acrometrix HBV Panel, Acrometrix, Menica, CA, USA) was also extracted for each run with the same protocol and1:10 diluted (from 50,000,000IU/mL to50IU/mL). For amplification, we used a primers/probe set designed under the auspices of “Agence Nationale de Recherches sur le SIDA et les hépatites virales” (ANRS 12187 project) and targeting a conserved region in the HBVS gene (nucleotide (nt) positions, 379–426). All runs were performed in a 50-μl volume containing DNA extract (10 μl), Master Mix (Platinum UGD, USA) (25 μl), pure water (HyClone Pure Water, Thermo Fisher Scientific, Waltham, MAUSA)(12.5 μl), forward primHBV1 (5’-GTGTCTGCGGCGTT TTATCA-3’) and reverse primHBV2 (5’-AGGCATAGCAGCAGGAT GAA-3’) primers at 10 μM (1μl each) and probe (5’FAM-TGCGGCGTTTTATCAT-MGB3’) at 5μM (0.5 μl). Each reaction consisted of: 2min at50°C and 10min at95°C; followed by 50 cycles of 15sec at 95°C and 1 min at 60°C each. The lower limit of quantification (LLOQ) of our technique was 100IU/mL and the lower limit of detection (LLOD) was 50IU/mL. Bivigou-Mboumba B, Amougou-Atsama M, Zoa-Assoumou S, Kamdem HM, Nzengui-Nzengui GF, Ndojyi-Mbiguino A, Njouom R, François-Souquière S (2018) Hepatitis B infection among HIV infected individuals in Gabon: Occult hepatitis B enhances HBV DNA prevalence. PLoS ONE 13(1): e0190592. doi: 10.1371/journal.pone.0190592 Unité Mixte de Recherche VIH et Maladies Infectieuses Associées (UMR-VIH-MIA), Libreville, Gabon, Unité Mixte de Recherche VIH et Maladies Infectieuses Associées (UMR-VIH-MIA), Libreville, Gabon, Laboratoire de Rétrovirologie, Centre International de Recherches Médicales de Franceville (CIRMF), Franceville, Gabon, Centre de Traitement Ambulatoire (CTA), Franceville, Gabon, Laboratoire de Rétrovirologie, Centre International de Recherches Médicales de Franceville (CIRMF), Service de Virologie, Centre Pasteur du Cameroun, Yaoundé, Cameroun, Laboratoire de Virologie, AP-HP, Hôpital Saint Louis; INSERM U941,Université Paris Diderot; Laboratoire associé au Centre national de Référence du VIH, Paris, France, Service de Virologie, Centre Pasteur du Cameroun, Yaoundé, Cameroun, Laboratoire de Rétrovirologie, Centre International de Recherches Médicales de Franceville (CIRMF) https://doi.org/10.1371/journal.pone.0190592 1 2018 Berthold Bivigou-Mboumba, Sandrine François-Souquière, Luc Deleplancque, Jeanne Sica, Augustin Mouinga-Ondémé, Marie Amougou-Atsama, Marie Laure Chaix, Richard Njouom, François Rouet 2018. HBV DNA Real Time Quantification according to ARNS 12187 project. protocols.io dx.doi.org/10.17504/protocols.io.kw4cxgw 2021-03-29 03:09:52
VBRC Base By Base: Switching between Mouse Modes
 
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Nick Tang 10.17504/protocols.io.ecpbavn VERVE Net, Upton-Lab Viral Bioinformatic Resource CentreProvide databases of viral genomic information. Please check the Organisms menu to see which viruses we support: we’re now focusing on large DNA viruses The VOCs (Virus Orthologous Clusters) database is at the heart of our system. The database links directly to integrated tools for comparative analyses. VOCs sorts genes into ortholog clusters (e.g. RNA polymerase) to simplify data retrieval. Provide easy access to the genes, gene families, and genomes of the different virus families. via a unique series of powerful Java tools that support multiple computer platforms (see VBRC Tools menu). design and build software to tackle specific bioinformatics/virology problems, often in collaboration with virologists. Rally the research community to provide expert curation of these viral genomes by: Adding value to GenBank sequences through enhancing and updating genome annotations Linking to research reviews/papers for the research community. Collaborate with researchers to help on specific bioinformatics problems, e.g. Custom searches of the databases Building new features into our tools Help with genome annotation University of Victoria: Department of Biochemistry and Microbiology http://athena.bioc.uvic.ca/ 1 2016 Nick Tang 2016. VBRC Base By Base: Switching between Mouse Modes. protocols.io dx.doi.org/10.17504/protocols.io.ecpbavn 2021-03-29 03:09:54
UC Davis - Running Wheel
 
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Jennifer Rutkowsky 10.17504/protocols.io.yv9fw96 Mouse Metabolic Phenotyping Centers Voluntary wheel running provides detailed information on the running capacity and activity patterns of control, treated or mutant mice. Running activity is evaluated from the number of rotations of a stationary wheel detected by sensors attached to running wheel. Running wheel activity provides information regarding when the animal runs, for how long and far, providing temporal and longitudinal data. This system may also be used as a means of providing voluntary exercise in metabolic or exercise based studies. University of California, Davis https://mmpc.org/shared/document.aspx?id=301&docType=Protocol 1 2019 Jennifer Rutkowsky 2019. UC Davis - Running Wheel. protocols.io dx.doi.org/10.17504/protocols.io.yv9fw96 2021-03-29 03:09:55
Isolation of temperate phages by plaque agar overlay
 
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John H. Paul and Markus Weinbauer 10.17504/protocols.io.dpd5i5 VERVE Net This protocol gives a method for isolating temperate phages from marine viral concentrations. This protocol uses the conventional plaque agar overlay and looking for turbid or haloed plaques, a hallmark of temperate phages.Paul, J. H., and M. Weinbauer. 2010. Detection of lysogeny in marine environments, p. 30–33. In S. W. Wilhelm, M. G. Weinbauer, and C. A. Suttle [eds.], Manual of Aquatic Viral Ecology. ASLO.Please see the full chapter for additional details. Manual of Aquatic Viral Ecology http://www.aslo.org/books/mave/MAVE_030.pdf 1 2016 John H. Paul and Markus Weinbauer 2016. Isolation of temperate phages by plaque agar overlay. protocols.io dx.doi.org/10.17504/protocols.io.dpd5i5 2021-03-29 03:09:54
Nuclei Isolation for SNARE-seq2
 
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Carter Palmer, Christine Liu, Jerold Chun 10.17504/protocols.io.8tvhwn6 BICCN This protocol is intended to be used for the isolation of nuclei from fresh-frozen brain tissue in preparation for analysis by Single-Nucleus Chromatin Accessibility and mRNA Expression sequencing (SNARE-seq). It has been applied to tissues from mouse, marmoset, and human. University of California, San Diego, and Sanford Burnham Prebys Medical Discovery Institute, University of California, San Diego, and Sanford Burnham Prebys Medical Discovery Institute, Sanford Burnham Prebys Medical Discovery Institute 1 2020 Carter Palmer, Christine Liu, Jerold Chun 2020. Nuclei Isolation for SNARE-seq2 . protocols.io dx.doi.org/10.17504/protocols.io.8tvhwn6 2021-03-29 03:09:55
Collection of FOCUS™ Mitochondria Kit protocols
 
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G-Biosciences 10.17504/protocols.io.e8tbhwn G-Biosciences https://www.gbiosciences.com/image/pdfs/protocol/786-022_protocol.pdf 1 2016 G-Biosciences 2016. Collection of FOCUS™ Mitochondria Kit protocols. protocols.io dx.doi.org/10.17504/protocols.io.e8tbhwn 2021-03-29 03:09:52
Protein Coomassie Blue Staining
 
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Alan Cone 10.17504/protocols.io.fw8bphw Ju Lab Wright State University 1 2016 Alan Cone 2016. Protein Coomassie Blue Staining. protocols.io dx.doi.org/10.17504/protocols.io.fw8bphw 2021-03-29 03:09:52

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