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We narrowed to 968 results for: LAT;

Showing: 541 - 560 of 968 results

  1. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...2024). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https:/...primer binding site and a reverse transcription template (RTT). The design of the pegRNA has been shown...editing for precise and highly versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. ...

  2. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...Thursday, or, at the latest on Friday. It’s best to split cells before the plates are more than 80-85%...Benjamin Franklin)! It’s good practice to label your plates and/or flasks with your name, date, the cell line...disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied phycology27.6...

  3. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known...

  4. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ...of a 5-6 day old human blastocyst [6]. They are isolated mostly from abnormal and/or supernumerary preimplantation...Characteristic ESC ASC Self-renewal High Low once isolated from the human body Usage Universal. Can generate...concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation rejection...when used therapeutically High Low (if ASCs are isolated from the patient) For many years, ESCs or ASCs...

  5. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac cells....propagated through multiple cells to do things like translate perceived signals to physical actions. Understanding...serotype specificity during delivery (more on this later). Chemigenetic sensors take a hybrid approach and...JF). The voltage-dependent changes in rhodopsin modulate the degree of quenching of the fluor via FRET....

  6. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology...full-color images of proteins. But they are still flat. To truly appreciate the 3D structure of these proteins...   He then said “I hadn’t realized how perfectly flat the complex is on this surface”….where the complex...cleavage”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting...

  7. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    ...protein or its role in a disease, researchers often isolate proteins of interest and examine them using biochemical...protein, it's important to validate its function relative to the untagged, wild-type protein. You may, for...the protein of interest and then analyzing the relative colocalization of their fluorescent signals and...One should also be aware of additional problems related to the properties of FPs themselves. These can ...

  8. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...institutions recently reported their inability to recapitulate the results of Gao et al 2016 in a letter to...NgAgo DNA guide is inexpensive and can be 5’-phosphorylated in house using a simple PNK reaction or can... the instability of the supposedly alien 5’-phosphorylated ssDNA guides, which might be rapidly degraded... an ssDNA guide. It is possible that, during translation, only a small fraction of nascent NgAgo protein...

  9. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...double-stranded break. HDR copies the sequence from a repair template with flanking sequence homology for error-free...systems, and NHEJ will be favored even if a repair template is present. HDR also presents a cloning dilemma...CRISPR gene editing. In organisms where HDR is downregulated, MMEJ represents another method for making targeted...’s become clear that the power of this editing platform lies in the diversity of nucleases and their applications...

  10. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ... up after trying out the new protocols.io beta platform for sharing, annotating, and storing life science...molecular cloning protocols, including making LB agar plates, restriction digest of plasmid DNA, and bacterial...bacterial transformation. And more plasmid-related educational resources can be found in our Plasmids 101 blog...the repository. These protocols show up on the related plasmid pages and can help scientists use these...

  11. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

  12. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...field to a grinding stop and only now, 2 decades later, is it finally back in the limelight. On December...and severe visual impairment that faithfully recapitulated the human condition. Luck brought all these ...X-linked retinitis pigmentosa, achromatopsia, and age-related macular degeneration are ongoing and some will ...capsid variants, designing expression cassettes to regulate and optimize transgene levels, and large scale...

  13. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...would convert adenine to inosine, which the translational machinery treats like guanine. This RNA editor...would permit point mutations in RNA which could recapitulate or rescue known pathogenic alleles, or introduce...ADAR will preferentially edit an adenine if the template has a cytosine mismatch at that position. PspCas13b-ADAR2DD...RNA Editing Capabilities through RESCUE Two years later, the Zhang lab expanded the RNA editing toolkit ...

  14. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ...trivia, and everything in between). Eleven years later,  Addgene is a company of fifty instead of three...situation- do we work so hard so we have time to play later? Or, does our play encourage us to work hard? Whatever...employees. The teambuilding culture helps us build relationships and thereby work better together.I realize I...encourage everyone to make an effort to build relationships with their colleagues. It can be easy to get...

  15. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...bi-directionality. For predominately anterograde tracing, biotinylated dextran amines (Veenman et al., 1992) or fluorescent...., 2007; plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have...overview see Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size which makes them capable...Transport: Cargo-Specific Mechanisms of Motility and Regulation. Neuron 84(2), 292–309. PubMed PMID: 25374356...Veenman, C.L., Reiner, A., and Honig, M.G. (1992). Biotinylated dextran as an anterograde tracer for single-...

  16. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ...available to many countries, import and export regulations, differing customs processes, and the distances...through the use of popular Chinese social media platforms.  As you can imagine, shipping to so many countries... a Primer Creating Bacterial Glycerol Stocks Inoculating a Liquid Bacteria Culture Collections ... Primers for PCR Agarose Gel Electrophoresis Inoculating Liquid Bacterial Cultures Restriction Digest ...

  17. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...called CROPseq-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing...fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also led to similar...donor site; SAS, splice acceptor site; pA, polyadenylation signal. Image reused from Riedmayr et al. 2023...therapies. REVeRT supports advanced gene expression regulation, enhancing both the mechanisms of delivery and...

  18. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...Possible roadblocks only become obvious at rather late stages (i.e. postdoc level) and can include the ...non-academic career options require additional study for late career scientists to become more appealing on the.... The issue was with a custom-made reagent to manipulate cells (manufactured by one of the larger biotech...signals are obtained. Or someone may heat an ELISA plate during incubation steps in order to enhance the ...

  19. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...binding specificity: SpCas9 variant Mutations (relative to SpCas9) PAM sequence D1135E variant ...bacterial species Many more Cas9 orthologs have been isolated, and our understanding of their PAMs is shown ...same species. Expanding the CRISPR toolbox The isolation of novel CRISPR proteins has and will continue... an intrinsically high-fidelity genome-editing platform. Genome Biol 19: . https://doi.org/10.1186/s13059...

  20. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ...to be translated into Modern Greek to be easily accessible today. Roughly speaking, we translated our software...sprints is themed and the stories in that sprint relate to the theme. For example, we could have a shipping...feature to work. We will then go back to this area later and tidy it up. Conclusion While none of us are...
Showing: 541 - 560 of 968 results