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We narrowed to 527 results for: HA tag

Showing: 441 - 460 of 527 results

  1. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ... disrupting ribosome-mRNA complexes. Recent work has adapted this technique to monitor local translation...contains 24 copies of SunTag, a synthetic scaffold that can recruit GFP fused to the SunTag-specific antibody...Imaging by Coat protein Knock-off) instead takes advantage of the ribosome’s movement. When the ribosome ...indicating translational repression. During later stages, single RFP-labeled osk and Oskar protein were ...scFv. Each time the reporter is translated, the SunTag sites on the nascent polypeptide recruit scFV-GFP..., as the authors of these papers did. Since the SunTag-based system tethers mRNAs to the plasma membrane...Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the Brightest...

  2. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ... AAV quality control Historically, AAV vector QC has been sparse including only titration of AAV particles... used SMRT sequencing, TA-based ligation and tagmentation-based methods for AAV genome sequencing (Tai...self-complementary AAVs and the TA-based ligation and tagmentation preparation approaches described previously ...single-stranded and both TA-based ligation and tagmentation require a double-stranded template.  Using viral...the mix to serve as effective substrates for tagmentation-based library preparation procedures. This finding...packaging plasmids. We decided to use this to our advantage by designing a Python script that looks for pre-defined...

  3. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...native organism.” This is possibly a better idea and has been used successfully in the past (Angov et al.,...functionally analyze the protein produced to ensure that it has folded properly. You may be able to avoid getting...normally found at low abundance in E. coli. The advantage of producing additional tRNAs is that you can ...

  4. How to Write a Scientific Review Article

    Type
    Blog Post
    ...requires. Using the (Last name, Year) format also has the benefit of exposing you to relevant researchers...just gotta do it." At the end of the day, someone has to write the article and that someone is going to...retreat to the woods for months at a time, so take advantage of whatever time you have. If you’re like me, ...

  5. Management for Scientists: Managing vs. Leading

    Type
    Blog Post
    ...leadership. Perhaps one of your advisors or mentors has urged you to work on developing your “leadership ...just have to know it when you see it? “The Three Stages of Your Career: Doer, Manager, Leader” by Charlotte...importance of these very different roles. At any stage in our careers we may be in a cycle of doing, managing...

  6. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...in state machine technology? Biological research has produced a massive amount of information regarding...two-input gene regulation programs. The software has two components - a database of pre-compiled register...reporter-based) each come with their advantages and disadvantages. Sanger sequencing is simple and the...

  7. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...possible angle to leave no stone unturned, but cancer has been producing landslide after landslide, only adding...used immunotherapeutic agents. Currently the FDA has 12 approved monoclonal antibodies targeting the PD-L1...bacteria, the tumor cells that escape the elimination stage acquire genetic alterations and alter cell-surface..., by way of favorably attaining the equilibrium stage. This can be called “stalemate equilibrium” where...the acute immune responses that eliminate early stage cancer cells, chronic inflammatory responses can...

  9. Antibodies 101: Flow Cytometry Controls

    Type
    Blog Post
    ...rather than for setting up your initial voltages and gates. Ashlyn has a research background in CD8+ T cells...color controls are used to help you set up your voltages and can also be helpful for compensation. You ... cells. This can be achieved by adjusting your voltage using your single color controls (Figure 1A), instead... using beads and not experimental samples, the voltages and compensation you set from your single color...

  10. An Introduction to Adenovirus

    Type
    Blog Post
    ... virulence (Tollefson et al., 1996) – cell death has been reported when using vectors, and immune inactivation...the infection to sometimes result in cell death, has led to interest in using adenoviral vectors for vaccine...death protein (E3-11.6K) is required at very late stages of infection for efficient cell lysis and release...

  11. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...high for both plasmids, so each plasmid “thinks” it has a higher copy number than it actually does, and neither...large, low copy virulence plasmids. This strategy has been successful in displacing, or “curing” the virulence...Using plasmid incompatibility for therapeutic advantage Although it may seem like plasmid incompatibility...

  12. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...FLP efficient The first version of FLP discovered has a temperature optimum of 30 °C and is therefore inefficient...evolution led to the identification of FLPe which has a temperature optimum of 37 °C (Buchholz, Angrand...Properties of FLP Recombinase Evolved by Cycling Mutagenesis.” Nature biotechnology 16(7): 657–62. PubMed ...

  13. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...particles, the functional infection unit. Each particle has a viral envelope, which fuses with the target cell...the fear of random genomic integration causing mutagenesis in the host. Adenoviruses can deliver relatively...non-integrating and infect non-dividing cells. AAVs primary advantage over adenovirus is their low immunogenicity, which...Dividing Cells Moderate Immunogenicity Insertional Mutagenesis Possible Lentivirus Infects non-dividing...non-dividing cells Insertional Mutagenesis Possible Adenovirus Remains Episomal (Little Risk of...of Insertional Mutagenesis) High Gene Expression Infects Non-Dividing Cells Large Packaging Capacity (... AAV Remains Episomal (Little Risk of Mutagenesis) Very Low Immunogenicity Infects Non-Dividing...

  14. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...applicable in DSB-induced genome engineering, it has biological importance for the repair of breaks that...directed repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made... chromosomal aberrations. Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination...

  15. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...combining the function of both editors into one? This has been done by fusing the adenine and cytosine editing...deaminases with a Cas9 nickase (Zhang et al., 2020). It has increased CBE activity and reduced RNA off-targeting...engineering and directed evolution of the TadA domain has yielded base editors that are equally efficient at...which uses a different mechanism than base editing, has allowed scientists to replace entire sequences of...properties of the individual Cas9. They also mutagenized the cytidine deaminase portion of the base editor...

  16. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...bridge helix. Created with BioRender.com.   Cas9 has two major lobes, called NUC (for nuclease) and REC...gray). Target DNA strand, light gray.   Once it has found a PAM site, Cas9 needs to check for complementarity...orange) and HNH (red) domains, while the REC lobe has been hidden for visibility. A RuvC loop caps the ... and different techniques all bring their own advantages—and blind spots—to these questions. Each new ...

  17. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...emerging validation technology. Digital PCR has several advantages over mismatch nuclease assays that will...As mentioned, these assays have several major advantages over the commonly employed mismatch nuclease ...bi-allelic mutations: Perhaps the most noteworthy advantage of digital PCR assays is that they are easily .... 6. Fu, Y. et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells...

  18. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ...senior mentors available to go around. My advice has been to find 5 interested peers and form your own...step-by-step, how-to guide to get one started. Advantages of the group mentoring format Adding peer advice...Because the participants are at a similar development stage, they have a lot to learn from each other. Some ...the group. The group can be one gender and career stage (e.g. all women, all men, all postdocs, all grad...support each other as you transition to the next stage in your careers! Stay in touch…this group will be...

  20. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...the most widely used CRISPR nuclease, recent work has shown that alternative enzymes may better suit specific...Joung and Jennifer Doudna Labs used targeted mutagenesis to generate hyper accurate Cas9 (HypaCas9). This... sites. Using a combined random and directed mutagenesis approach, the Anna Cereseto Lab created pX-evoCas9...experimental approaches, knowledge of both the advantages and limitations of each Cas enzyme can help determine...
Showing: 441 - 460 of 527 results