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We narrowed to 954 results for: TES

Showing: 521 - 540 of 954 results
  1. How to Keep a Lab Notebook for Bioinformatic Analyses

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    Blog Post
    ...together. I also prefer the number system to using dates as labels, since I often work on multiple projects...sure that the original code remains in tact! Take notes Comment in your scripts: Comments on what a given...who are still learning the basics. However, these notes take up a lot of space and don’t tell you why you...
  2. Screening for Successful Genome Editing with Digital PCR

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    Blog Post
    ...calculated (details in [2]) and can be used to quantify rates of mutation in a bulk or single cell-derived genomic...instructions on how to design such assays, in addition to testing their performance (2). Remember that although digital...transfection of a novel TAL effector nuclease (TALEN) facilitates efficient knockout of HIV co-receptor CCR5. Nucleic...
  3. Plasmids 101: Screening Strategies Used in Plasmid Cloning

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    Blog Post
    ...below). Plating the transformation on ampicillin plates containing X-gal allows you to identify cells that...that when a portion of the gene is deleted, it creates a non-functional 𝛽-galactosidase. Complementing...this deleted portion, for example on a plasmid, creates a functional 𝛽-galactosidase. Blue-white screens...
  4. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

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    Blog Post
    ...can rapidly and efficiently evaluate Cas9 cutting sites and activity (Knudsen et al., 2018).  To create ...lab compared editing at three different genomic sites in the AAVS1 locus using GEmCherry2. In this experiment...again that sgRNA structure and genomic recognition sites play a large role in CRISPR-Cas9 efficiency. They...
  5. Giving gRNAs a Facelift - Synthetic and Beyond

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    Blog Post
    ...repeat region on its 3’ end. This repeat region associates with the trans-activating crRNA (tracrRNA) which...at the same site, is 2’-fluoro (2’-F), which substitutes a fluorine for the hydroxyl group at the 2’ position...coupling several of them together, for example phosphorothioates with 2’-O-methyl modifications, proved more...
  6. Hot Plasmids - May 2022

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    Blog Post
    ... both on and off target. Across all loci tested, error rates overall ranged from 0.3% to 10%, in comparison...Importantly, this is true in both mice and nonhuman primates (for AAV.CAP-B10, for example).  Given that the...
  7. CasPEDIA: A Functional Classification of Cas Enzymes

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    Blog Post
    ... trans activity. AsCas12a, on the other hand, generates staggered DNA nicks at its target in cis and cleaves... used by Jinek et al. 2012, or browsing for the latest AsCas12a plasmids, Addgene and CasPEDIA have got...from CasPEDIA.   How can I use it? CasPEDIA operates like many search engines, featuring both a general...
  8. Flow Cytometry Readouts: Yes, No, and Everything in Between

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    Blog Post
    ...to the right indicates a higher signal, or upregulation, and a peak to the left indicates a lower signal...continue gating populations of interest with normal gates, just based on a histogram plot instead of a dot...
  9. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    Blog Post
    ... packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus using different...post‐transcriptional regulatory element; sequence that stimulates the expression of transgenes via increased nuclear...U5 and the rest of the provirus. 3’ LTR Terminates transcription started by 5' LTR by the addition...
  10. Antibodies 101: Flow Cytometry Controls

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    Blog Post
    ...companies or check the literature. Pro tip! Early gates, like your live/dead markers or commonly used markers...specific cell populations, often used in the later gates, are more likely to require FMOs. Isotypes Isotype...rather than for setting up your initial voltages and gates. Ashlyn has a research background in CD8+ T cells...
  11. Viral Vectors 101: Viral Applications

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    Blog Post
    ...be used to express proteins. Viral expression cassettes typically contain a promoter (inducible or constitutive...lineage, and so much more! Libraries allow you to test thousands of elements simultaneously in your system...dose. Vaccine development strategies. Image courtesy of Gorry et. al, 2007.   Clinical applications...
  12. 3 Challenges in Plant Synthetic Biology

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    Blog Post
    ...infancy. A long timeline from transformation to testing and a lack of well-characterized genetic tools ...excited to see how this exchange of genetic tools facilitates plant science. Challenge #3: Intellectual property...in 2004, she accepted a commission in the United States Air Force. She left active duty to pursue a degree...
  13. Antibody Guide

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    Guide
    ...response has developed, the antibodies are collected, tested against the antigen, packaged, and sold. This is...immortalized B cells). The genetic reorganization that creates variety in the antibody allowing for their extreme...class therefore has a valency, or number of binding sites available per antibody unit, which is typically ...multiplied by two, as each antibody has two binding sites. Isotype Description Configuration and Valency Subclasses...Monomer with a valency of 2 IgE Protects against parasites and is responsible for driving allergic reactions...in the lab. Check for and record any expiration dates. Buffers Buffers are a concern for conjugating and...reaction in the presence of several different substrates. The luminescence produced by the reaction can...
  14. Plan Your Experiment

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    Guide
    ...additional sites throughout the genome. These sites are called off-targets. Be aware of off-target sites when...efficiency. So while testing multiple gRNAs is a good idea for most CRISPR experiments, testing multiple pegRNA...efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing. Scientific...that there could be downstream translation start sites that could allow for expression of a truncated protein...PAM sequences or Cas9 enzymes with flexible PAM sites . Base Editing : Target the site of your desired...complement to your target sequence with no homologous sites elsewhere in the genome. In reality, a given gRNA...genetic cargo (in this case, CRISPR components) integrates into the genome. Lentiviral vectors are a good...
  15. Seven Tips for Using LinkedIn as a Scientist

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    Blog Post
    ...keep track of how you met the connection, as these notes are saved in the messaging stream. Unfortunately...LinkedIn is not the platform for politics, family updates, or click bait of any kind. For more on inappropriate...However, do comment on other people’s news and updates. Share your own good ideas, events and favorite...
  16. Troubleshooting Your Plasmid Cloning Experiment

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    Blog Post
    ...antibiotic in your plate might not work. Check the plates! Streak on them an E. coli  strain known to be ...growing fine, it is probably time to prepare new plates with a fresh stock of antibiotic. Many small colonies... different temperature (30°C) for the growth on plates and in liquid culture. If you don’t get any positive...
  17. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

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    Blog Post
    ...caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle progression and enhances tumor... TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells proliferate... 2004). In fact, Sanford H. Barsky and his team tested the influence myoepithelial cells had on the conversion...
  18. Choosing the B(right)est Fluorescent Protein: Photostability

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    Blog Post
    ... oxygen concentration may affect photobleaching rates (Shaner et al, 2008). This brings us to the key ...application (Goedhart et al, 2012). The third issue relates to the environment of the fluorescent protein. ...The issues with quantification of photobleaching rates can be summarized as follows: Instead of determining...
Showing: 521 - 540 of 954 results