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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
Protocol for predicting specialist herbivore distribution from host distribution data
 
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Joshua Fuller 10.17504/protocols.io.9gvh3w6 A necessary, but difficult, part of ecology and conservations is to try to create order and meaning from chaos. An example of such a task is the need to predict where an organism may exist at any given point in a landscape. In the past, this was done by systematically surveying the entire landscape, a lengthy and expensive endeavor. With the current pace of habitat loss and climate change, new methods were created to efficiently determine the distribution of a species using computers and new statistical algorithms. These new methods still require people in the field taking GPS points of the target species. A way to ease the workload on researchers in the field is to use data generated through citizen science projects such as iNaturalist. Citizen scientists can rapidly generate a tremendous amount of species GPS points by just exploring parks and backyards. That data can be directly utilized by researchers to create a species distribution model (SDM). The goal of this protocol is to take that a step further by generating an SDM for a plant, Tsuga canadensis, and then using that model to determine the distribution of an invasive specialist, Adelges tsugae, on the plant. By the end of this protocol, you should be able to create an SDM, generate a map using GIS software, and critically evaluate the results of the model to determine any conclusions that can be drawn from the model. University of North Carolina at Chapel Hill 1 2020 Joshua Fuller 2020. Protocol for predicting specialist herbivore distribution from host distribution data. protocols.io dx.doi.org/10.17504/protocols.io.9gvh3w6 2021-03-29 03:09:18
Guava Flow Cytometer - Troubleshooting "Tray Hold Off State"
 
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Steven Wilhelm 10.17504/protocols.io.iv9ce96 The Aquatic Microbial Ecology Research Group - AMERG (The Buchan, Zinser and Wilhelm labs), Samantha R Coy's Protocols Please contact Dr. Steven Wilhelm ([email protected]) for additional information regarding this protocol. 1 2017 Steven Wilhelm 2017. Guava Flow Cytometer - Troubleshooting "Tray Hold Off State". protocols.io dx.doi.org/10.17504/protocols.io.iv9ce96 2021-03-29 03:09:18
An error-corrected panel-based next-generation sequencing assay for ultra-sensitive somatic mutation detection in ctDNA
 
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Heidi Fettke 10.17504/protocols.io.bb3siqne Circulating tumour DNA (ctDNA), fragmented DNA shed into the circulation from tumour cells, holds the potential to become a minimally-invasive biopsy material with which to interrogate the tumour landscape. However, ctDNA can be highly diluted in normal cell-free DNA (cfDNA) (released into the blood primarily by hematopoietic cells), and thus robust liquid biopsy assays with high analytical sensitivity are required for ctDNA to reach its full clinical potential in molecular genotyping.This protocol uses optimised off-the-shelf reagents in order to generate indexed and molecularly-tagged cfDNA and germline DNA libraries compatible with Illumina short-read sequencing. This protocol involves the hybridisation-capture of targeted regions of the genome using RNA baits - and has been used previously with a 42-gene panel 447 kbp in size. Also described here is the bioinformatic tools used for data processing and variant identification. The combination of deep-sequencing and error-correction based on molecular barcoding allows high sensitivity and specificity to be generated at low variant frequencies. Department of Medicine, School of Clinical Sciences, Monash University, Australia 1 2020 Heidi Fettke 2020. An error-corrected panel-based next-generation sequencing assay for ultra-sensitive somatic mutation detection in ctDNA. protocols.io dx.doi.org/10.17504/protocols.io.bb3siqne 2021-03-29 03:09:18
Preparation of soil bacteria for FCM
 
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Laura Espina 10.17504/protocols.io.bh8qj9vw Cardiff University 1 2020 Laura Espina 2020. Preparation of soil bacteria for FCM. protocols.io dx.doi.org/10.17504/protocols.io.bh8qj9vw 2021-03-29 03:09:18
Complete Sample Protocol for Measuring IC50 of Inhibitor PD168393 in A431 Cells Responding to Epidermal Growth Factor
 
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LI-COR Biosciences 10.17504/protocols.io.grxbv7n LI-COR Biosciences Developed for: Aerius, Odyssey® Classic, Odyssey CLx, and Odyssey Sa Infrared Imaging Systems LI-COR Biosciences, Lincoln, Nebraska USA https://www.licor.com/documents/ebj3m4pfmhv3tzq5ivhzkyrzk8f3i7z3 1 2018 LI-COR Biosciences 2018. Complete Sample Protocol for Measuring IC50 of Inhibitor PD168393 in A431 Cells Responding to Epidermal Growth Factor. protocols.io dx.doi.org/10.17504/protocols.io.grxbv7n 2021-03-29 03:09:15
Drug sensitivity assay (CCK8)
 
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Shuangshuang Li 10.17504/protocols.io.h4mb8u6 Li S, Jin X, Wu H, Wang Y, Li X, Guo Y, Liang S, HA117 endows HL60 cells with a stem-like signature by inhibiting the degradation of DNMT1 via its ability to down-regulate expression of the GGL domain of RGS6. PLoS ONE 12(6). doi: 10.1371/journal.pone.0180142 https://doi.org/10.1371/journal.pone.0180142 1 2018 Shuangshuang Li 2018. Drug sensitivity assay (CCK8). protocols.io dx.doi.org/10.17504/protocols.io.h4mb8u6 2021-03-29 03:09:15
Anti-Neu5Gc Antibody Kit Protocol - Western Blot
 
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Sam Li 10.17504/protocols.io.97hh9j6 BioLegend The Anti-Neu5Gc Antibody Kit contains the essential monospecific polyclonal chicken IgY antibody, along with a negative control primary antibody to detect the presence of Neu5Gc on glycoconjugates by Western blot (WB). Samples to be evaluated are first subjected to SDS-PAGE, followed by transfer to a nitrocellulose or polyvinylidenedifluoride (PVDF) membrane. The membrane is then incubated with affinity-purified polyclonal anti-Neu5Gc to determine the presence of Neu5Gc on the protein of interest.The antibody provided in this kit has been shown to identify as little as 5pmol of Neu5Gc per µg glycoprotein, which is at or below the current detection limit for conventional analysis by acid release, purification, DMB derivatization, HPLC, and electrospray mass-spectrometry. The Western blot provides additional information in that it confirms that Neu5Gc is directly linked to the glycoprotein of interest rather than to an accompanying sample component. BioLegend https://www.biolegend.com/protocols/anti-neu5gc-antibody-kit-protocol-western-blot/4300/ 3 2019 Sam Li 2019. Anti-Neu5Gc Antibody Kit Protocol - Western Blot. protocols.io dx.doi.org/10.17504/protocols.io.97hh9j6 2021-03-29 03:09:18
Visualisation of bacteria around roots
 
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Andreea S 10.17504/protocols.io.bjzpkp5n iGEM Groningen 2020 This protocol is carried out to observe the growth of the bacterial biofilm along with the roots in the rhizosphere. In the paper by Bennett, R. and Lynch, J (1981), they inoculated the plant with the bacterial strain and analyzed the growth of the bacterial biofilm around roots. University of Groningen 1 2020 Andreea S 2020. Visualisation of bacteria around roots. protocols.io dx.doi.org/10.17504/protocols.io.bjzpkp5n 2021-03-29 03:09:15
In vitro transcription of guide RNAs and 5'-triphosphate removal
 
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Mark Dewitt, Julia Wong, Beeke Wienert 10.17504/protocols.io.bfxfjpjn CornLab sgRNA template assembly, in vitro T7 transcription, and sgRNA column cleanup to remove 5'-triphosphate groups UC BERKELEY-IGI, UC BERKELEY-IGI, UC BERKELEY-IGI https://doi.org/10.1371/journal.pbio.2005840 11 2020 Mark Dewitt, Julia Wong, Beeke Wienert 2020. In vitro transcription of guide RNAs and 5'-triphosphate removal. protocols.io dx.doi.org/10.17504/protocols.io.bfxfjpjn 2021-03-29 03:09:15
Serum Biochemical Indexes Detection
 
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Honghao Zhao, Jasmine Chong, Rong Tang, Li Li, Jianguo Xia, Dapeng Li 10.17504/protocols.io.smeec3e This protocol is widely used in serum biochemical indexes detection, it provides an rapid and reliable technique for obtaining relative concentrations of multiple blood biochemical indices simultaneously. College of Fisheries, Hubei Provincial Engineering Laboratory for Pond Aquaculture, National Demonstration Center for Experimental Aquaculture Education, Huazhong Agricultural University, Wuhan 430070, China, Institute of Parasitology, and Department of Animal Science, McGill University, Saint-Anne-de-Bellevue, QC H9X 3V9, Canada, College of Fisheries, Hubei Provincial Engineering Laboratory for Pond Aquaculture, National Demonstration Center for Experimental Aquaculture Education, Huazhong Agricultural University, Wuhan 430070, China, College of Fisheries, Hubei Provincial Engineering Laboratory for Pond Aquaculture, National Demonstration Center for Experimental Aquaculture Education, Huazhong Agricultural University, Wuhan 430070, China, Institute of Parasitology, and Department of Animal Science, McGill University, Saint-Anne-de-Bellevue, QC H9X 3V9, Canada, College of Fisheries, Hubei Provincial Engineering Laboratory for Pond Aquaculture, National Demonstration Center for Experimental Aquaculture Education, Huazhong Agricultural University, Wuhan 430070, China 1 2018 Honghao Zhao, Jasmine Chong, Rong Tang, Li Li, Jianguo Xia, Dapeng Li 2018. Serum Biochemical Indexes Detection. protocols.io dx.doi.org/10.17504/protocols.io.smeec3e 2021-03-29 03:09:15
Immunohistochemical staining of heparan sulfate (HS) and collagen type XVIII (col18) core proteins in islet beta cells of formalin-fixed human pancreas and isolated human islets
 
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Lora Starrs, Debra Brown, Sarah Popp, Andrew Ziolkowski, Charmaine Simeonovic 10.17504/protocols.io.bkqykvxw Paraffin sections (4 μm thickness) of formalin-fixed human pancreases and isolated human islets were treated with 0.05% pronase for antigen retrieval. HS and Col18 HSPG core proteins were detected immunohistochemically using 10E4 anti-HS (US Biological/Amsbio) and anti-Col18 (Santa Cruz), respectively, with horseradish peroxidase-conjugated rabbit anti-mouse Ig (Dako). Background staining was checked using the corresponding isotype control Ig instead of the primary antibody. 3-amino-9-ethylcarbazole (AEC) was used as the chromogen. For morphometry, stained sections were imaged using a light microscope with attached camera (Olympus BX41). Image J software with color deconvolution plugin was used for the quantitative analysis of the % of islet area stained The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University, The John Curtin School of Medical Research, The Australian National University https://doi.org/10.1371/journal.pone.0191360 2 2020 Lora Starrs, Debra Brown, Sarah Popp, Andrew Ziolkowski, Charmaine Simeonovic 2020. Immunohistochemical staining of heparan sulfate (HS) and collagen type XVIII (col18) core proteins in islet beta cells of formalin-fixed human pancreas and isolated human islets. protocols.io dx.doi.org/10.17504/protocols.io.bkqykvxw 2021-03-29 03:09:15
COVID-19 SCAN molecular workflow
 
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Lea Starita 10.17504/protocols.io.bebkjakw Coronavirus Method Development Community Molecular Detection of SARS-CoV-2 for SCAN the Greater Seattle Coronavirus Assessment Network .justify:after { content: ""; display:inline-block; width: 100%; } .justify:after { content: ""; display:inline-block; width: 100%; } The SCAN real-time RT-PCR assay for SARS-CoV-2 contains primer/probe sets that target the Orf1b and S genes, designed against the SARS-CoV-2 genome (GenBank: MN908947.3). The primer/probe sets were analyzed in silico for specificity for this specific betacoronavirus by Thermo Fisher. All assays are performed on total nucleic acids extracted from upper respiratory tract swabs, including mid-turbinate and nasopharyngeal (NP) swabs stored in Universal Transport Media. The assay limit of detection of 5.6 (Orf1b) and 12.9 (S) molecules per reaction was determined from serial dilutions of synthetic DNA of the target sequences. These probes reproducibly detected SARS-CoV-2 RNA from cultured virus from USA-WA-1/2020 (BEI resources/ATCC) and from 21 specimens confirmed to be positive by the Washington State Department of health. Specificity was determined by lack of detection of SARS-CoV-2 from specimens that are known to be positive for seasonal coronavirus strains HKU1/NL63 or 229E/OC43 and/or other respiratory viruses. These specimens are further inferred to be negative by the fact that they were obtained in the greater Seattle area prior to the first known reports of SARS-CoV-2 in China. .justify:after { content: ""; display:inline-block; width: 100%; } .justify:after { content: ""; display:inline-block; width: 100%; } University of Washington 1 2020 Lea Starita 2020. COVID-19 SCAN molecular workflow. protocols.io dx.doi.org/10.17504/protocols.io.bebkjakw 2021-03-29 03:09:15
PCTE Protocol for VLP enumeration
 
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Ken Stedman, Geoffrey Diemer, Jennifer E Kyle 10.17504/protocols.io.eaubaew VERVE Net An alternative epifluorescence microscopy (EFM) technique for virus enumeration has been developed using polycarbonate Track Etch (PCTE) filter membranes in place of aluminum oxide Anodisc membranes. The established EFM technique, first developed by Hennes and Suttle in 1995 and later expanded upon by Noble and Fuhrman in 1998, uses supported 20 nm pore-size Anodisc filter membranes to determine virus abundance in natural environments. Increased price and sporadic availability of Anodisc filters stimulated the evaluation of alternative filters for use in the procedure. The feasibility of using 30 nm pore-size PCTE filters for virus enumeration was assessed using the Anodisc filter procedure as a control. Although virion particle counts are slightly less precise using PCTE filters, they offer a substitute for Anodiscs while requiring only minor adjustments to the established protocol. Per slide, the PCTE costs approximately ten times less to prepare than the Anodisc-based method. Stedman Lab, Stedman Lab, Stedman Lab http://web.pdx.edu/~kstedman/PCTE_virus_counting_protocol.pdf 1 2016 Ken Stedman, Geoffrey Diemer, Jennifer E Kyle 2016. PCTE Protocol for VLP enumeration. protocols.io dx.doi.org/10.17504/protocols.io.eaubaew 2021-03-29 03:09:16
Fungal extraction from Xylosandrus compactus
 
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James Skelton, Craig Bateman, Jiri Hulcr 10.17504/protocols.io.bnuumeww Protocols Bark Beetle Mycobiome This protocol describes how to extract ambrosia fungi from Xylosandrus compactus, the black twig borer. This protocol is part of the Bark Beetle Mycobiome (BBM) Research Coordination Network. For more information on the BBM international network: Hulcr J, Barnes I, De Beer ZW, Duong TA, Gazis R, Johnson AJ, Jusino MA, Kasson MT, Li Y, Lynch S, Mayers C, Musvuugwa T, Roets F, Seltmann KC, Six D, Vanderpool D, & Villari C. 2020. Bark beetle mycobiome: collaboratively defined research priorities on a widespread insect-fungus symbiosis. Symbiosis 81: 101–113 https://doi.org/10.1007/s13199-020-00686-9. University of Florida, University of Florida, University of Florida 1 2020 James Skelton, Craig Bateman, Jiri Hulcr 2020. Fungal extraction from Xylosandrus compactus. protocols.io dx.doi.org/10.17504/protocols.io.bnuumeww 2021-03-29 03:09:16
FlowCam Standard Operating Procedure
 
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Jacob Harris 10.17504/protocols.io.f2tbqen Protist Research to Optimize Tools in Genetics (PROT-G), Alverson Lab The FlowCam can be used to take quick cell density measurements of liquid samples.  2 2016 Jacob Harris 2016. FlowCam Standard Operating Procedure. protocols.io dx.doi.org/10.17504/protocols.io.f2tbqen 2021-03-29 03:09:17
Colonization of aposymbiotic Aiptasia with Symbiodinium
 
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Grossman Lab, Pringle Lab 10.17504/protocols.io.rh4d38w Aiptasia-Symbiodiniaceae Model System This protocol describes briefly how to perform colonization of aposymbiotic Aiptasia with Symbiodinium cultures.The method is based on the one described in Xiang et al., 2013. Carnegie Institution for Science - Department of Plant Biology, Stanford University 1 2018 Grossman Lab, Pringle Lab 2018. Colonization of aposymbiotic Aiptasia with Symbiodinium. protocols.io dx.doi.org/10.17504/protocols.io.rh4d38w 2021-03-29 03:09:16
QuikChange Mutagenesis
 
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James Fraser 10.17504/protocols.io.dij4cm Fraser Lab Fraser Lab, UCSF 1 2015 James Fraser 2015. QuikChange Mutagenesis. protocols.io dx.doi.org/10.17504/protocols.io.dij4cm 2021-03-29 03:09:09
PCR Prep from cDNA for IVT Reactions
 
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Allen Institute for Brain Science 10.17504/protocols.io.bcsfiwbn BICCN, Allen Institute for Brain Science This protocol describes PCR1 and PCR2 reactions beginning with cDNA to generate transcript templates ready for IVT reaction. This protocol is written for 192 (2x96) reactions but can also be used for fewer or more reactions by adjusting the amount of materials and equipment appropriately. Allen Institute 1 2020 Allen Institute for Brain Science 2020. PCR Prep from cDNA for IVT Reactions. protocols.io dx.doi.org/10.17504/protocols.io.bcsfiwbn 2021-03-29 03:09:09
Laboratory calibration of soil moisture sensors in porous media (repacked soils)
 
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Soham Adla, Neeraj Rai, K Sri Harsha, Shivam Tripathi, Markus Disse, Saket Pande 10.17504/protocols.io.pjidkke Technische Universität München, Kritsnam Technologies, Kritsnam Technologies, Indian Institute of Technology, Kanpur, Technische Universität München, Delft University of Technology 1 2018 Soham Adla, Neeraj Rai, K Sri Harsha, Shivam Tripathi, Markus Disse, Saket Pande 2018. Laboratory calibration of soil moisture sensors in porous media (repacked soils). protocols.io dx.doi.org/10.17504/protocols.io.pjidkke 2021-03-29 03:09:09
PCR-based assay for genotyping of the slick mutation in cattle
 
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Elizabeth Jannaman, M. Sofia Ortega, Tad S. Sonstegard, Peter Hansen 10.17504/protocols.io.6kxhcxn University of Florida Bovine Embryo Lab Several mutations exist in the prolactin receptor gene (PRLR) of cattle that cause a short, sleek haircoat.. The mutation is dominant so slick-haired cattle can have either one ortwo copies of the mutation. The assay described here detects the most-well studied mutation (SLICK1)that causes translation of a truncated PRLR of 461 amino acids. Originally described in Senepol cattle,this mutation has also been introduced into Holsteins in Puerto Rico, Florida and elsewhere. Note thatthe assay will not detect other mutations in PRLR that also cause the slick phenotype. University of Florida , University of Missouri - Columbia, Recombinetics, Inc., University of Florida https://animal.ifas.ufl.edu/hansen/lab_protocol_docs/SLICK.pdf 1 2019 Elizabeth Jannaman, M. Sofia Ortega, Tad S. Sonstegard, Peter Hansen 2019. PCR-based assay for genotyping of the slick mutation in cattle. protocols.io dx.doi.org/10.17504/protocols.io.6kxhcxn 2021-03-29 03:09:09

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