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We narrowed to 1,011 results for: CAL;

Showing: 621 - 640 of 1011 results

  1. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...protein expression in mammalian cells: easy to use biological tools are abundant! In this post, we will review...that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 fused to a transcriptional repressor... protein, then these systems need to be site-specifically knocked in. This can be labor intensive and ...and target-specific protein degradation. Nature Chemical Biology. May 2018. 14(5): 431-441. PubMed PMID...

  2. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    ... Not only is CRISPR having effects on various biological disciplines, the base technology itself is constantly...at non-target sites. Means to decrease these so-called “off-target effects” have included using a Cas9... the Broad Institute, the Joung lab at Harvard Medical School, and the Doudna Lab at Berkeley set out ...afforded by gRNA binding at non-target sites would theoretically decrease off-target cutting. Figure 1...

  3. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...researchers to identify and isolate very specific biological targets. However, as popular and useful antibodies...antibodies, creating a potentially unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively...has specific binding and has a strong signal. We call the unique pattern of the antibody binding “distinct...TS (2015) The Economics of Reproducibility in Preclinical Research. PLoS Biol 13(6): e1002165. https://...

  4. A Look at Addgene's QC Process

    Type
    Blog Post
    ...plasmids. These are prone to error — both human and biological. Even the most high-fidelity polymerases can ...the correct inserts, tags, promoters, and other critical functional components. Fun fact! Some older plasmids...sequences. These sequences can also be confirmed or theoretical. No matter the level of information that is provided...the raw NGS reads and reassemble using a program called Geneious. Here, the raw reads are mapped against...

  5. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    ...choosing antibodies   Complexity is a key factor Biological systems tend to be complex with a variety of ...multiple targets in parallel. In these cases, it is critical that users choose antibodies that are easily distinguishable...staining multiple targets indirectly, you must strategically plan primary/secondary antibody pairing such...Fisher, S. K. (1993). Chapter 15 Postembedding Immunocytochemical Techniques for Light and Electron Microscopy...

  6. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ... Jesse Zalatan’s lab deposited plasmids for a chemically inducible CRISPRa system in yeast is mediated...Find the yeast CRISPRa plasmids here Reduce the scale of your CRISPR screens by using this lentiviral ...paired guides per construct and allows a reduced scale in screens without any reduction in performance ... channelrhodopsin (C1V1) targeting parvalbumin cortical interneurons is now available as a ready to use...

  7. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    ...things more interesting, we are working in a new technical discipline that nobody understands (synthetic ...thing is opening a pizza joint. Silly ideas (mining local salt deposits to sell at farmers markets), and serious...that survived the literature review, survived technical feasibility analysis, and was a marketable product...and rapidly indicate their presence. They are genetically modified organisms, and GMOs have a bad reputation...

  8. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    ...therapeutic strategies to target it, than you typically would at a microbiology meeting. Create an outline...lab mates, and PI can provide feedback on your technical presentation, but those outside of your field ... through the list at the session so you can strategically use your time. Don’t have access to a list of...titles to find the ones you’d like to revisit. Strategically traverse the sea of posters If you come prepared...

  9. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ... Liu, 2023). This method uses a prime editor (typically a Cas9 H840A nickase fused to a reverse transcriptase... design of the pegRNA has been shown to play a critical role in the editing efficiency of prime editing...experimentalist, and especially shines once large-scale prime editing experiments come into play. Nicolas...s41587-023-01678-y Li, X., Chen, W., Martin, B. K., Calderon, D., Lee, C., Choi, J., Chardon, F. M., McDiarmid...

  10. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...your gene of interest is difficult to target specifically with a gRNA). Be sure to keep these techniques...sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes advantage...homology is required, but 6 to 14 kb of homology is typical for targeting constructs. In the example shown ...cells participating in development or other physiopathological events to which the chosen promoter responds...

  11. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...can be long—5 years or more, especially in the biological sciences—whereas industry postdocs have a set...course, you might have less time to explore scientifically since you can’t afford to spend too much time...Unless your academic lab is closely involved with clinical trials or has ties to industry, the drug discovery...industry postdoc, you might find yourself feeling scientifically isolated if no other postdocs are in your department...

  12. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...margins of my protocol. While these tips weren’t critical to the experiment, they definitely made my life...qPCR Protocol for more details. Pro Tips When calculating the titer for your AAV, remember to account for...control (NTC) should always be included. While the physical titer generated by qPCR is useful, I test a few...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...

  13. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ... 2017. At Addgene, we continually use the Basic Local Alignment Search Tool (BLAST) provided by NCBI. ...BLAST sequence databases from the dropdown menu. Typically, we use the default nucleotide database “Nucleotide...” program for blastn so that you can extract practically every single reliable base pair from your results... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...

  14. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...-length viral genome (Figure 2). While it is technically possible to encode all four viral genes in the... (see Uses of Minigenome System). From a more technical standpoint, a plasmid with all four genes and ... gene by altering the amount transfected (see Technical Tips). Minigenome replication and transcription...compounds which often target the viral polymerase. Technical tips It is important that all of these plasmids...

  15. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...lab tool. In the classic two hybrid system (also called the interaction trap), genes encoding proteins ...experiments. False positives can occur even in small scale experiments, although these can often be accounted...that the configuration of your fusion proteins physically blocks the binding sites for the protein partner...Tachihara, K., Umezawa, Y.l. Split luciferase as an optical probe for detecting protein-protein interactions...

  16. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...that, they also help us keep track of the tubes’ physical location in the lab. These barcodes additionally...  freezers depending on the sample type. It is critical for us to know which plate, rack, and freezer ...performs an average of 140 transformations. The lab typically transforms newly deposited samples from Monday...Excel sheet.3. Be specific  - When assigning a physical location to your samples, include as much detail...

  17. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...been working on development of technologies for biochemical analysis of genome functions such as transcription...researchers to use enChIP in a wide variety of biological systems. We are currently developing several ...Therefore, my lab is dedicated to development of new biological technologies. The locus-specific ChIP technologies... effort to interact with each other, which is critically important, especially for young researchers. ...

  18. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...studying parenting strategies and make important ecological indicators since species live in water and on... as the only Monarch butterfly food. They have chemical pathways generating interesting drug-like compounds...Lab (@EvoEmbryo) September 24, 2018   Xenopus tropicalis (Gary Gorbsky) This is not a new experimental...plants and are unusual enough to be interesting biologically. They have economic importance as harmful weeds...

  19. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ... defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base...precisely changes a single base with an efficiency typically ranging from 25-75%, while the success of precise... in the DNA by cutting only a single strand, dramatically reducing the likelihood of detrimental indel...known 'DNA adenosine deaminases' which would theoretically give rise to an adenine base editor. A massive...

  20. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    ...gives Snapgene great exposure to the community of biological scientists who use Addgene everyday. To get this...from other institutions personally on conference calls and make sure to maintain these relationships by...speakers for a meetup or recruit volunteers for your local animal shelter. There are many opportunities so ...Universities themselves can help you do this through practical tools like alumni networks. I find it useful to...
Showing: 621 - 640 of 1011 results