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We narrowed to 856 results for: HA

Showing: 221 - 240 of 856 results

  1. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...While much of CRISPR research has focused on genome editing, numerous discoveries have been made using...cleave the DNA. Much of the research using dCas9 has focused on transcriptional activation using a fusion...CRISPR-mediated purification of a specific genomic region has also recently been adapted to isolate genomic loci...tag systems including Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification...for many other CRISPR techniques, this method also has great potential for improvements and adaptations ...

  2. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...altering the sequence of the protein. As DNA synthesis has become quite cost-effective it seems easy for scientists...optimize your gene of interest? Codon optimization has become extremely useful in expressing functional ...neurons in vivo (Slimko and Lester, 2003).  Addgene has a large selection of codon optimized genes, so check...which have even been linked to some diseases. It has been reported that 5-10% of human genes contain a...//doi.org/10.1093/nar/15.3.1281 Slimko EM, Lester HA (2003) Codon optimization of Caenorhabditis elegans...

  3. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...Cas9, but the ever-expanding repertoire of CRISPR has brought many other Cas effectors to light. One lesser-known...effector, and type IV with an unknown effector. Type I has subtypes A through G, with different compositions... Thanks to this complex, developing Cas3 tools has been challenging. It can be difficult to ensure comparable... job — Cas see DNA, Cas chop DNA. Cas3, however, has two jobs, one as a nuclease and one as a helicase...easier to work with than Cas3. Future targets Cas3 has the potential to be incredibly useful in therapeutics...

  4. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...decades, recombinant Adeno-associated virus (AAV) has been widely used to deliver genetic tools to the ...The most famous ones are influenza hemagglutinin (HA), myelocytomatosis viral oncogene (myc), simian virus...limitations of epitope tags and fluorescent proteins, it has been crucial to develop new tracers that enable accurate...are known to be notoriously difficult to label. It has also been shown that dendrites in the stratum oriens... provided better labels than fusions to 3 or more HA tags. 4. High-resolutions microscopy As indicated...

  5. The Challenges of Cell Culture

    Type
    Blog Post
    ...Addgene's Special Collections). Arguably, oncology has benefited most as cancer cells are amenable to culture...misidentified cell lines. The solution Fortunately it has never been easier to conduct high quality cell culture...identity. For example, perhaps a working cell bank has been made by a research institution (an excellent...at the scene of a crime (every human in the world has a unique STR profile) and can also be used as a paternity...topics. The LGC Group also provides standards and has laboratories to carry our independent quality control...

  6. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...firm The path to a career as a Grant Professional has been a long and winding path for many in the field...?  In the past 5-10 years, the Grants Profession has become just that, a profession. A grant professional...and the increasing regulatory process for grants has fostered the demand for experts in the field of grants... find your people Find someone who is working or has worked as a Grant Professional to mentor and coach...includes the social media community GrantChat. She has served in the nonprofit and government sector for...

  7. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ... in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes...protein interactions in living cells. Since then FRET has also been used to measure conformational changes,...brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP. The stoichiometry...Fluorescence lifetime imaging microscopy method FLIM has been developed more recently and is the most rigorous...emergence of faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...

  8. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    ...policy. Since early March, the open access publisher has required authors to include a note as to where readers...how much data was necessary and reasonable. PLOS has since clarified their data sharing policy and recently...Who is to say that the initial data-generating lab has efficiently trawled their own data? A distinct (perhaps...many have multiple, complex panels. PLOS, itself, has not finalized its definition of necessary data requirements.... Secondly, science is highly technical and this has a natural, albeit unintentional, tendency to work...

  9. Meet Aavery, Blugene's New Virus Friend!

    Type
    Blog Post
    ... and polling, Blugene's new virus friend finally has a name! We thank you for your participation in suggesting...play on viral vector terms. And, the name Kendall has a special place in our hearts: our first two offices...

  10. Scientist Networking: What is an Informational Interview?

    Type
    Blog Post
    ...Training as a scientist in the academic system has many pluses. I delighted in my graduate school years...call this building relationships because networking has gotten a bad reputation (as in…”I just hate networking...connections. A mistake made by many novice networkers has a potential to alienate connections. If meeting with...

  11. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...targeting. Fast forward a decade, and the CRISPR field has exploded with dozens of Cas enzymes and variants ...scientific community. What are the features? CasPEDIA has collected relevant information on all the Cas enzymes...cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the other hand, generates...learn about Cas domains. CasPEDIA either directly has this information or links out to this content and...

  12. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ...ElizabethKenkel) December 5, 2018 How social media has helped me in science It may have taken a bit of FOMO...Here are three examples of how using social media has helped me or my lab mates: For me, using social ...with social media as a scientist Using social media has enriched my professional development, and while it...helped you fit social media into your busy schedule. Has social media helped you advance your research or ...

  13. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    ...being targeted via RNAi. Recently the Vellai lab has created an improved RNAi vector that leads to significantly...sequence of the toehold switch. For example, Green has also developed toehold switches to detect Zika virus...protein motifs than previously thought. Sidhu’s group has made several hundred SH3 domain containing plasmids.... In addition to these constructs, the Sidhu lab has made hundreds of additional plasmids available through...

  14. Science Guides

    Type
    Guide
    ... Science Guides Addgene has created the following guides to help you start learning...

  15. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    ...and post-docs alike are wondering where the time has gone. Are we any closer to graduating, publishing...unsure of the next steps. Harvard PI Angela DePace has put personal development at the center of her lab... and postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine...

  16. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ... field of induced pluripotent stem cells (iPSCs) has been around for 10 years. In that time, scientists... of Myc or other oncogenes after differentiation has been linked to tumor formation in iPSC-derived and...iPSCs compared to other delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic...factors driven by one promoter. While lentivirus has similar rates of reprogramming when compared to MMLV-derived...mechanism between a vector and chromosomal DNA. PB has a cloning capacity of ~9-14 kb. This system is transfected...cell division. A drawback to this approach is it has lower levels of efficiency at generating iPSCs, usually...reprogram cells is that it’s simple and efficient. It has a high efficiency of reprogramming when compared ...

  17. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...wildtype Cas9 with optimized chimeric gRNA has high efficiency but has been shown to have off-target effects... the Genome Engineering Using CRISPR/Cas Systems has expanded out of control. To further consolidate these... a new system, developed by the Zhang lab, which has comparable efficiency to the optimized chimeric design...sequence is present in both alleles. The Zhang lab has found that if a CRISPR is used to engineer a locus...relative to where the homology ends? A9: The Zhang lab has always used homology arms that are less than 15 bp...recombination? A10: The most we've tried to insert so far has been 1 kb. We used homology arms that were 800 bp...for positive ones.  -- OR -- If your HR template has a selection marker such as puromycin, you can (also...

  18. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...to find the perfect combination. The EcoFlex Kit has a wide array of DNA parts that can be assembled into...yeast MoClo is also for eukaryotes! The technology has been used to optimize protein secretion and expression...possible through MoClo. The MoClo Plant Parts Kit has everything you would expect in a MoClo kit plus the... general plant cloning experiments. Addgene also has two additional MoClo plant kits: Plant Parts Kit ...; Chamness et al., 2023). The Plant Parts II kit has a focus on infrastructure with shuttle vectors, several... gene silencing vectors. The Plant Parts III kit has a focus on transformation and genome engineering....Brychcy et al., 2023). For plant biologists, Addgene has not one, not two, but three CRISPR MoClo kits for...

  19. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ... 16, 2020 by Beth Kenkel.  CRISPR genome editing has quickly become a popular system for in vitro and ...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room for additional...the number of targetable loci, and truncated Cas9 has much lower efficiency than its wild-type counterpart...amino acids, can also be packaged into AAV. NmeCas9 has the added advantage that it can be turned off by ...daunting, it’s actually a challenge that the field has overcome with split AAVs. In general split AAVs break...

  20. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ... maintain itself within host cells.  The plasmid has multiple ori that are functionally differentiated...various hosts (Pagatto and Dillon 2001).  The plasmid has its own replication initiation factors and proteins...the host. Host range depends on whether the host has the components to recognize the ori and allow for...restriction enzymes (Meyer et al. 1977). So if a plasmid has few restriction sites, it may be able to sneak past...commonly used bacteria in lab manipulations. Table 1 has a detailed list of the bacterial species these plasmids...
Showing: 221 - 240 of 856 results