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Showing: 561 - 580 of 955 results
  1. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...BioID, BirA* is fused to a bait protein and it biotinylates proteins that are within a ~10 nm radius of ...the bait proteins were tagged with. It’s best to test all combinations and pick the conditions that give...interactome. Further characterization of these candidates is critical to understanding how they interact...
  2. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ... experiments is cutting efficiency so that the greatest effect possible will be achieved. On the other... an NG PAM.   Cas PAM sequence  Additional notes SpCas9 NGG - Cas12a TTTV Great for AT-rich...coverage in the human genome. SpRY Cas9 has PAM sites covering the majority of the genome! Want to...
  3. Hot Plasmids February 2024

    Type
    Blog Post
    ...co-incubate it with an “assist peptide” that facilitates endosomal escape. Combining Cas9-PAGE with lentiviral... in rodents have not been widely successful in primates. The authors used a multispecies approach to generate...endothelial-specific tropism in rodents and broad tropism in primates. Nature Communications, 14(1), 3345. https://doi.org...
  4. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ... high complexity Golden Gate Assemblies. Image courtesy of New England Biolabs.   DAD Tools The three...input a sequence, SplitSet finds optimal fusion sites to split it into multiple fragments. You can define...sequences or set a narrow search window for a site—or sites—that must be used. Each of the tools works with ...
  5. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ... plate format yielding anywhere from two to six plates of 96 samples per week. This is where our new process... process only takes about one day, even for six plates (576 individual plasmids)! These libraries are ...takes about two days to complete. After the run completes, we begin our assembly process. Again, thanks ...
  6. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ... used to integrate the tagged AID* into genomic sites of interests via CRISPR techniques. This tool-kit...activated, Cas9 cleavage can induce DSBs within minutes. New CRISPR plasmids for Borrelia burgdorferi from...are now available. These libraries encode target sites flanked by randomized nucleotides in place of a ...
  7. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ... real challenge is finding relevant papers. The latest installment of the Early Career Researcher Toolbox...around since 1996, but is getting an update! The latest version can be found here. The new PubMed will ...cite the most), and who the author influences (who cites the author the most). Best uses Finding papers frequently...
  8. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    ...identified and interpreted in less than a couple of minutes. How does one accomplish all of that? What follows...The UCLA Library offers some Powerpoint based templates with different column numbers for a good starting...starting point. Adobe also offers free poster templates, although most are vertical.  Large central box...
  9. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...vectors carrying fluorescent markers are used as it eliminates the need for antibody staining. Check Out Our...multiplicity of infection (MOI) to ensure only one copy integrates, selecting by antibiotic-resistance or single...
  10. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ... (green or blue) within a population of timer indicates the age of the pool: the higher the ratio, the...differentiation revealed by temporal cell labeling." Diabetes 58.8 (2009): 1863-1868. PubMed PMID: 19478145....programmable synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-...
  11. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ... learn what motivates them (keep in mind that it can be very different from what motivates you!), know...moment while reviewing the results of my Myers-Briggs test:  my undergrads were different people! More specifically...
  12. Tips to Make the Most of a Scientific Conference

    Type
    Blog Post
    ...you’d like to go to. During the conference Take notes during talks. If you have questions during the talk...poster.” Have a notebook handy so you can jot down notes or questions from your poster viewers and trade ...whenever you have a small break), write down some notes on the card so you remember who they are and what...
  13. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...AAV8 for kidney cells — can help improve infection rates in your experiment. Figure 2: Visual representation...important to note, however, that while AAV rarely integrates into the genome, CRISPR may increase this rate..., nor is it a global occurrence, as integration sites seem to only occur in the target cells or tissues...
  14. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...career path long ago. Use your analytical mind to test and discover what career suits you best I began ... make a trophy for the winner. We promoted the contest for about two months, and we got one submission...girl about fossils. After we talked for about ten minutes, she said, “Boy, I love science,” and I realized...
  15. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ... the expression system you choose in many ways dictates the best method for introducing those CRISPR components...will want to invest some time in optimizing and testing the efficiency of CRISPR delivery. There are a ... at Addgene that have been published as CRISPR testing tools: Traffic Light Reporter System: Can evaluate...
  16. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...endonuclease gene within its own cut site1. In heterozygotes, the endonuclease cuts the wild-type chromosome...Both methods are easy to implement and have been tested in yeast5. Ecological confinement involves performing...area where escaped organisms won't be able to find mates2. For example, ongoing experiments attempting to...
  17. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...detection of the product. As an added benefit, replicates are built into the technology. One well (containing...Have a dedicated bench with a dedicated set of pipettes for ddPCR set-up. Prepare the master mix in a ...low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has been ...
  18. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own proteins, making the purification... the efficacy of our antibodies through stress testing. An aliquot of our recombinant antibodies is stored...aliquots frozen immediately post-harvest. Stress-tested antibodies that are not equally effective are shipped...
  19. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...to ~4-10x linearly! This expansion physically separates molecules while maintaining their relative positions...understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux, et al. 2021; Gambarotto, ...new insights into the cytoskeleton of malaria parasites including the conservation of a conoid. PLoS Biology...
Showing: 561 - 580 of 955 results