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We narrowed to 956 results for: tes

Showing: 521 - 540 of 956 results
  1. Antibody Guide

    Type
    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...
  2. Plan Your Experiment

    Type
    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...
  3. Screening for Successful Genome Editing with Digital PCR

    Type
    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...
  4. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    ... start, the team inserted AsLOV2 into conserved sites in all eight surface-exposed loops of an mCherry...SOS), a guanine nucleotide exchange factor that activates Ras. They expressed this OptoNB-SOS fusion protein...membrane-localized mCherry. With darkness, SOS binds Ras and initiates the extracellular signaling-regulated kinase (...
  5. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    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...
  6. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    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...
  7. Antibodies 101: Flow Cytometry Controls

    Type
    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...
  8. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    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...
  9. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    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...
  10. Hot Plasmids - May 2022

    Type
    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...
  11. Viral Vectors 101: Viral Applications

    Type
    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. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    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...
  13. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    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...
  14. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    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...
  15. 3 Challenges in Plant Synthetic Biology

    Type
    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...
  16. Lentiviral Vector Guide

    Type
    Guide
    ...incompetent by using four plasmids instead of three and eliminates the requirement for Tat. Always includes deletion... transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods...target species. Although chromatin availability facilitates integration, it does not explain the lentiviral...more accurately predict and understand integration sites have led to the development of multiple genome-wide...al. (2020) for examples). Pseudotyping Tropism dictates which types of host cells the lentiviral vector...glycoprotein — transduce airway epithelium and cardiomyocytes in utero Rabies glycoprotein — transduce neural...for genome-wide mapping of lentiviral integration sites. Molecular Therapy — Methods & Clinical Development...
  17. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    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...
  18. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ... cross-talk with the other orthologs. Ma et al. tested their dCas9 variants using gRNAs specific for telomeric...individual colors to 6. A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a...separate vector. Figure 2: CRISPRainbow generates 6 unique colors and a white composite. gRNAs with...
  19. Seven Tips for Using LinkedIn as a Scientist

    Type
    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...
Showing: 521 - 540 of 956 results