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We narrowed to 960 results for: REP

Showing: 401 - 420 of 960 results

  1. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...jejuni (Cj) NNNNRYAC Streptococcus thermophilus (St) NNAGAAW Treponema denticola (Td) NAAAAC...in mammalian cells. The REPAIR (RNA Editing for Programmable A to I Replacement) system fuses Cas13b to...when trying to edit a gene using homology directed repair, since HDR-mediated gene editing is most efficient...Hu et al., 2018).  In 2020, the Kleinstiver lab reported the development of near-PAMless Cas9 variants....below.  Cas9 species PAM sequence ’) Streptococcus pyogenes (Sp) 3' NGG Staphylococcus aureus...

  2. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...different CGBE tools allow the researcher to direct the repair of the abasic site. But what if an editor could...excision is the most common type of base excision repair. But mammals also have an enzyme to remove inosine...collaborative team from the David Liu and Dali Li labs reported a similar tool shortly thereafter, using mouse... or to create the substrate for a base excision repair pathway. The team behind AYBEv3 introduced the ... the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template —...editors Excises G, T, or C, respectively, to be replaced with another base. References Chen, L., Hong...Transversion Editors Read about four base editor reporters Resources on Addgene.org Browse CRISPR plasmids...

  3. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...express my concerns and request a replacement. I got my replacement in the end and the issue was resolved... A degree in the life sciences prepares one for numerous non-academic careers. Still, many start their...support roles. Examples are Technical Service Representative, Technical Support Specialist, Technical Support...less suitable for troubleshooting or issuing replacement products. In order to find good and helpful answers...certainly there won’t be a shortage of amusing reports where someone tries to “optimize” a particular ...job. Solutions may also be achieved by simply replacing the product in question. This can solve the problem...

  4. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...amplify and design primers that will bind to and replicate it. The following image shows the ends of the ...mutation than restriction enzyme based cloning. DNA replication by PCR has error rates that range from roughly...

  5. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...gradients? Excellent! Now, let’s head west, intrepid reader! Protein Prep The first step in any western blot is...minutes) at 37 °C, if you suspect aggregation in your prep. If your proteins are the wrong size on your gel...will simply load an extra ladder or technical replicate (ie, leftover sample) to fill the relevant wells...antibody’s dynamic range. A) The protein dilutions, representing expected concentrations, are all within the ...antibody’s dynamic range. B) The protein dilutions, representing expected concentrations, are outside of the ...blot, you may want to consider using a different reporter or, if you don’t need to compare the protein amounts... amounts directly, stripping and reprobing. Just make sure to go from least to most abundant protein if...

  6. Save Time and Money by Making Your Own Competent Cells

    Type
    Blog Post
    ...reference: OpenWetWare Preparing chemically competent cells (protocol)  Preparing electrocompetent cells... Untergasser Lab Preparation of chemically competent cells (protocol) Preparation of electrocompetent...Research to make competent cells because cells prepared using this kit can be transformed without heat...of electrocompetent cells (protocol)  Cells prepared using the Z-competent TM buffer can be easily ...

  7. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ...fluorophore-conjugated streptavidin protein, which binds the primary antibody through the biotin-streptavidin interaction...detection. The advantage of this system is that streptavidin conjugates are commercially available for a ...fluorophore selection for panel design. Find streptavidin plasmids and biotin plasmids at Addgene!   ... two-protein labeling by utilizing(a) biotin-streptavidin binding and (b) antibody host species interactions...include transcription factors, histones, and DNA repair enzymes. Note that when using the nuclear fix/perm...or protein-protein interactions, like biotin-streptavidin binding and antibody host species reactivity...

  8. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    ...talk to the occasion Don’t just plan to reuse a prepared talk unless it perfectly fits the occasion.  I...figure out how to make it suspenseful. 4. Start preparing your talk early - like today Don’t wait until ...for poor quality slides with sufficient time to prepare (and it does take time). For important talks, ask...loves a great animation. 8. Predict questions, prepare answers Speaking pros always have a few extra slides...come up with clear and concise answers. This preparation will help you avoid giving only yes or no answers...their phones. Speaking longer just makes you seem unprepared or arrogant. A bit short is always better than...when I then ask “and who has deposited to the repository?” When most or all of the hands go down it is...

  9. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    ... are overrepresented on Twitter Mathematical, life, and physical scientists are underrepresented. It’s...interested in getting started? Reply in the comments below or tweet us your reply & #scientistswhotweet. And...users. Biologists rank #8. Figure 1: Highly Represented Scientific Disciplines on Twitter. Twitter users...Follower Network of Scientists on Twitter. Each dot represents a scientist. Only scientists who follow each ...

  10. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...engineered not to cleave DNA and instead activates/represses gene expression by various mechanisms. The RNA...cleaving of DNA occurs, not for deadCas activator/repressor experiments where only localization of the Cas...will generate a ssDNA break which can easily be repaired by cells with a near zero likelihood of mutation...insertion of DNA by the NHEJ pathway during DNA repair of the break to allow for site-specific detection... these approaches is that NHEJ must perform the repair to allow for detection, not Homologous Recombination...non-cutters If your Cas experiment involves activation/repression or any other effect which does not depend on ... be an appropriate method for an activator or repressor Cas. Similarly, Cas ChIP-Seq would also identify...

  11. Hot Plasmids - October 2022

    Type
    Blog Post
    ...the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. Think...pitch a Hot Plasmids section (antibodies and viral preps welcome!) here. Here's what you'll find in this...Liu Z, et al. Cell. 2022.  Find JEDI-2p viral preps here (and here!)Find JEDI-2p plasmids here!  Liu...adapted from Govorunova et al. 2022.     HcKCRs represent a long-sought tool for researchers to control ...s41593-022-01094-6. (This work was also shared as a preprint: https://doi.org/10.1101/2021.09.17.460684.)   ...

  12. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...data repository for validation data on Addgene/CLOVER viral vectors.  Caspedia: A data repository for ... Calculator app for iPhone Reagent and Data Repositories These links can help you find experimental and...for Cas enzymes.  Open repository of CRISPR screens Cellosaurus: A knowledge resource for cell lines IBEX...IBEX Knowledge Base: A data repository for validation data on IBEX reagents.  mAb3D atlas validated antibodies...human, and marmoset brain IHC.  FPbase: A data repository for fluorescent proteins and their properties...

  13. ReFigure: Save Scientific Figures into Dashboards and Share Your Insights

    Type
    Blog Post
    ... ReFigure. Reading and exploration including replications and experiments resulting in “negative data”...performance as in this antibody ReFigure Analyzing replications as in this ReFigure about whether Natronobacterium...reagent performs well, or you might publish a replication experiment that you conducted as part of setting...traditional publications. By submitting your data to a repository (Figshare is currently supported by ReFigure ... specialized knowledge of which findings are reproducible. For an example, see our ReFigures on the therapeutic... interim research products (pre-registration, preprints, data, software, micropublications) in grant proposals...

  14. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    ...in the specific repositories (Addgene, WormBase, Arabidopsis.info). These repositories will ensure that...published work meets the accepted standards of reporting in the discipline and reduces the dissemination...information that would prevent the study to be replicated or omission from the literature. I think that...data on ... The language is ... The general representation of data is... The figures are ... In general...the data can be deposited in the public on-line repository (like GitHub, Zenodo, Figshare, Triad) while ...

  15. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ...efficiency. If this finding is replicated in larger studies, it will represent a key advantage over Cas9, ... RETRACTED AND FOLLOW UP STUDIES HAVE FAILED TO REPEAT THE RESULTS DISCUSSED BELOW Biologists are going...the question on everyone’s minds is: could NgAgo replace CRISPR? Such a drastic shift won’t happen overnight...NgAgo is also able to mediate homology-directed repair when a template is supplied. Find Plasmids from..., this property could be an advantage. However, repair after base excision would alter the target sequence...

  16. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    ...Being an entrepreneur is difficult; it requires an immense amount of self-knowledge and an unwavering...become something tangible.  When you think of entrepreneurship in the 21st century, who do you think of?  ...server space. But what if you want to be an entrepreneur in the health sciences?  Is this a different...with innovative ideas in all facets of the entrepreneurial process in order to develop those ideas into...lessons I learned while becoming a health science entrepreneur at 22. Start with your network You've spent ...leave people remembering your potential as an entrepreneur, not your credentials. If you have an idea, ...

  17. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ...ApoB-NanoLuc reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...Cell, 2019. PubMed PMID: 30639098. LipoGlo: A reporter for measuring ApoB bound lipoproteins Article ... and localization in vivo. The LipoGlo system represents a new, intriguing approach to expand our understanding...cells. These changes were larger than previously reported for other organisms, and the Gottschalk group ...

  18. Hot Plasmids - January 2023

    Type
    Blog Post
    ...the new plasmids, antibodies, and viral preps in the repository through our hot plasmids articles. Think...pitch a Hot Plasmids section (antibodies and viral preps welcome!) here. Here's what you'll find in this...ubiquitous promoter drives expression of an mCherry reporter.  After using mCherry for FACS sorting of the ... you the benefit of a polyclonal, with the reproducibility of a (recombinant) monoclonal. Another reason...observed high fluorescence resilience to these preparations. After crystalizing the hfYFP structure, the...

  19. Science Career Options

    Type
    Blog Post
    ...When preparing to graduate from university, many students are confronted with the question ‘what now?...Michael Koeris' Journey from Grad Student to Entrepreneur Science careers divided by the most pertinent...actively require these skills, including: Science Reporter Scientific Sales Epidemiologist Scientific Recruitment... actively use these skills include: Science Reporter Scientific Illustrator Research and Development...desirable in a range of roles, including: Science Reporter Technical Writer Statistician Bioinformatician...Michael Koeris' Journey from Grad Student to Entrepreneur Learn to Network and Make the Contacts for Your...

  20. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...to primate neocortex. Cell Reports, 34(13). https://doi.org/10.1016/j.celrep.2021.108754 Ravindra Kumar...Look it up in Addgene’s Data Hub and the CLOVER repository of published systemic AAVs to find commonly used...removes, not just reduces, empty viral capsids in your prep can help reduce the risk of an immune response. ...primate. Many of the systemic capsids in our repository are deposited by the Gradinaru lab. You can visit...LY6C1) Stanton et al., 2023 * indicates viral preps available. Mechanisms of crossing the BBB are indicated...commonly requested systemic capsid in the Addgene repository. Like its parent AAV-PHP-B, it can be produced...region VIII of AAV9-X1.1 to that of AAV1. AAV1-X1 reproduces AAV9-X1.1’s brain endothelial cell-specific tropism...
Showing: 401 - 420 of 960 results