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We narrowed to 993 results for: Des

Showing: 261 - 280 of 993 results

  1. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...created SRIRACCHA in 2017 to enrich and identify desired CRISPR mutants (Wen et al., 2018). SRIRACCHA uses...constructs for the reporter to work and to make the desired genome edit:  A donor plasmid that contains a ...OH-T and doxycycline. A gRNA that targets your desired gene (whose sequence is also found in the reporter...well. The ratio of RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using...integrated GFP reporter leaving you with just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene...DSB is repaired by NHEJ which depending on the nucleotides inserted or deleted, can result in an in-frame...

  2. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    ...educational resources — including blog posts, protocols, guides, and collections — is a lot of material to sift... launched earlier in the year, as well as our redesigned homepage and our new search tool, are the results...a few of the strategies we used in our software design. In each case, we leveraged the team’s knowledge...per resource category to expose as filters and designed the search engine to return only the filters that... bits of information for a given resource, and designed the search engine to weight search results differently...'s new search Read to learn more about why we redesigned our homepage and mascot ...

  3. Distribution to Industry

    Type
    Collection
    ...Plasmid Gene/Insert PI Kits Kit name Type PI Description High Complexity Golden Gate Assembly Standards...PDB-deposited), disulfide-free, mesophilic, and designed to be diverse in topology, size, and isoelectric...Pooled Libraries Pooled Library name Type PI Description HR700_TP53 Exon Mutation Libraries CRISPR Thorsten...Parkinson's Disease; For target validation and drug design Antibodies Addgene distributes ready-to-use recombinant...materials and intellectual property rights, and provides liability protection for material providers. Addgene's...

  4. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...assessed for the desired parameter. A droplet containing a cell that is positive for a desired parameter is...their observations. However, while flow cytometry provides analysis at the cellular level, it cannot be used...degree of scatter is captured by detectors and provides useful information about the cell. Light that ... traveling is called forward scatter (FSC) and provides information about the relative size of the cell...initially traveling is called side scatter (SSC) and provides information about the cell’s complexity. A cell...side scatter (SSC). Users can gate around their desired cell population for further analysis. In this example...

  5. Promega Plasmid Collection

    Type
    Collection
    ...Promega.com. ID Plasmid Description Industry Plasmid Technology Overview This collection includes specialized Promega...contains plasmids developed by Promega scientists and designed to streamline molecular and cellular biology experimental...support a range of applications. The technology includes NanoLuc, HaloTag, SmBiT, LgBiT, HiBiT, and more... a new window) is an energy transfer technique designed to measure molecular proximity in living cells...

  6. All in a Twist: dsRNA

    Type
    Blog Post
    ...into agriculture as species-specific pesticides and insecticides, an advancement over previous chemical...prone to degradation – those are the words that describe RNA! Double stranded? Well, that’s just for DNA...also known as RNAi, are short, dsRNA molecules designed to degrade mRNA transcripts. RNAi activates the...sequence is found (typically RNA that the RNAi is designed against), RISC can repress translation and/or ... and repetitive sequence. All of the R-loops described above play distinct roles in promoting cellular...

  7. dTAG - You're it!

    Type
    Blog Post
    ... molecule degraders” recruit the proteasome to destroy targeted proteins. We previously developed small...locus-specific knock-in of FKBP12F36V. Detailed protocols describing the use of these plasmids are available here....Nabet et al., Nature Chemical Biology additionally provides detailed information on cloning procedures used...degradation, and lead author on the recent paper describing the dTAG technology. Follow him on twitter @behnamnabet... 29581585. This reference contains the full description of the dTAG system. The dTAG system was employed...Conditional Mouse Models CRISPR 101: Cas9 Nickase Design and Homology Directed Repair Plasmids for endogenous...

  8. Reproducibility for Everyone: Lessons from an Open Science Collaboration

    Type
    Blog Post
    ... could participate. Despite the group not having a public website, all the slides, handouts, and resources...science or even to discuss potential solutions. Despite all the talk, not much was being done to address...practices could be an actionable way to tackle widespread irreproducibility. Luckily, there were other ...thinking about reproducibility and shared similar desires to take action. Thanks to Teytelman’s vast network...practices could be an actionable way to address the widespread issue of irreproducibility. Use your network...

  9. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    ...approaches to gender equality over the past several decades: “Fix the Numbers of Women” focuses on increasing...-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...using both sexes in basic research is excellent design), but they used all female stem cells—this was ...environmental factors. How to include sex in experimental design Recognizing the importance of analyzing sex, the... above, another excellent resource for research design is Kalpit Shah et al., “Do you know the sex of ...Corporations need to integrate these insights into product design. But eyes have been opened—and we cannot return...

  10. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    ...monitored over time by sequencing the barcodes in the population (all barcodes can be amplified using the same...with roughly 73 million semi-random, 30 bp DNA barcodes that are integrated into target cells' genomes...ClonTracer system, lentiviruses containing these barcodes infect the target cell population at low multiplicity...replicate populations, they found that similar barcodes consistently made up the majority of the resistant...resistant populations. In fact, 88% of the barcodes within the resistant populations were shared by at least...

  11. TALEN Guide

    Type
    Collection
    ...components of the array recognize individual nucleotides. For over ten years, zinc finger (ZF) arrays ...that came out in the spring of 2011, the groups describe a set of customized plasmids that can be used ... a TALEN kit through Addgene. This kit, first described in Nature Biotechnology in August, uses a serial...in Nature Biotechnology early in 2011 and they described their kit for assembling TAL effector arrays in...effector and TALEN kits here . References Efficient design and assembly of custom TALEN and other TAL effector-based...

  12. Lentiviral Prep Service

    Type
    Collection
    ... Guide to Plasmid Pooled Libraries . ID Name Description Genes/Insert Selection PI Human knockout pooled...2A-Blast-WPRE) Zhang Pooled Barcoding Libraries ID Name Description PI 115643 Morris CellTag Barcode Library Version...Version 1 of the Celltag library contains 19973 barcodes to combinatorially index cells for single-cell...Version 2 of the Celltag library contains 4934 barcodes to combinatorially index cells for single-cell...Version 3 of the Celltag library contains 5737 barcodes to combinatorially index cells for single-cell...

  13. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...repository that will help epigenetic researchers custom design their own engineering plasmid tools in about a ...cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows for the assembly... cells that make up the human body and G-baToN provides versibility in tracking and modulating the interactions... inhibitor response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression...interests via CRISPR techniques. This tool-kit includes an array of FPs as well as additional useful tags...libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM...

  14. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a 3’UTR database specifically...specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures, ...imaging processing. The combination of these methods provides unique opportunities to develop computational ...Addgene In addition to the wealth of open resources described above, there were many plasmid tools presented...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...continuously curated as we continue to receive plasmids designed for the worm so check back frequently for what...

  15. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...high quality DNA. Figure 1 illustrates the three grades of DNA you may need based on their relative quality...components (i.e. buffers, columns, or beads) are designed to work together — no mixing and matching!   ...bacterial culture. Plasmid purification kits are designed to accommodate specific volume ranges of culture...column-based miniprep kit for example. These kits are designed to handle ~1–5 mL of liquid culture. Using a higher.... To help combat this, some manufacturers have designed eco-friendly DNA purification kits by reducing...Milllipore Sigma’s GenElute™ kits were all specifically designed with environmental sustainability in mind (Table... kits from a variety of manufacturers. It also includes a rough comparison of list price between these...

  16. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts CRISPR is a useful tool...screening experiments, due to the relative ease of designing gRNAs and the ability to modify virtually any ...can be found in the following table, which also includes information about the type of genetic modification... represent all 4096 (4 6 ) combinations of 6 nucleotides. Chong Zhang E. coli Genome-wide Inhibition Library...2 40,964 Ingolia lab S cerevisiae CRISPRi v1 – barcodes 181005 161769 Inhibition Yeast Ingolia NA ~6 44,580...Knockout Human Mali 3rd 1 74 Perturb-seq Guide Barcodes (GBC) 85968 Barcode Human Weissman 3rd N/A N/A...

  17. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...insertion site, the corresponding gRNA might be undesirable due to potential off-target sites or low predicted..., resulting in lower overall genome frequency. Despite the progress we’ve made, there is still a need ...bacterial polymerase subunit ω. This construct is designed to activate transcription from the accessory plasmid... stringency and promoting survival of only the broadest-targeting variants. For the final round of evolution...spectinomycin resistance. xCas9 3.7 showed the broadest PAM range, recognizing NG, NNG, GAT, and CAA PAMs...scaffold. With an NGG PAM, xCas9 3.7-VPR constructs modestly outperformed SpCas9-VPR in activating transcription...

  18. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...genetic locus when complexed with an appropriately designed small RNA, or guide RNA (gRNA). The gRNA conventionally...exploited to either inactivate a gene or introduce a desired genetic alteration. Listen to Our Podcast Interview... truncated such that it exhibits 15 or fewer nucleotides of complementarity to a target site, Cas9’s DNA...truncated gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased.... An additional benefit of the use of truncated guides is that they can be delivered into systems that...Cas9 species. Within a single cell, full-length guides targeting one set of genetic loci can be introduced...

  19. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...amazing project. You’ve written your protocols, designed your experiments, and prepared your reagents. ...following cloning methodologies have been extensively described in other articles; so if you are interested in...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...-generated insert. When mixed, topoisomerase I provides both the endonuclease and ligase function while...restriction enzymes. In Golden Gate cloning, the desired end construct does not contain the type IIS recognition...This feature biases the reaction in favor of the desired assembly to make the process close to 100% efficient...

  20. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...degradation of the tagged protein. Degrons are destabilizing domains which will be identified by various ...proteasome for degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and...protein for imaging. Given its popularity and widespread usage, a proteolysis-targeting chimera (PROTAC... The auxin-inducible degron (AID) system is the oldest system reviewed here. The AID system relies on ...Thankfully, this is a hot topic with many resources! Bondeson et al. (2022) reviews all the tags discussed here...choice!   References and Resources References Bondeson, D.P., Mullin-Bernstein, Z., Oliver, S. et al....
Showing: 261 - 280 of 993 results