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  1. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ... GFP is such a well-validated, all-round good performing probe, it is the first choice when selecting ...reduces its size. The substantial size of GFP may interfere with the activity or the localization of the fusion...C-terminus of another protein. This may, however, interfere with the function of the protein. This is clearly...
  2. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...analysis is performed yielding a graph of edits in the input population or clone.   How to perform a TIDE...homology-directed repair events, or maybe you’re performing a single base pair edit with a prime editor. ...
  3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ... virus directly using saliva without having to perform RNA extraction but that may work only if there ... Motif) for target cleavage. This test can be performed in two steps for challenging samples as in Two-Step-SHERLOCK...fluorescently labelled substrate, gets activated and performs non-specific collateral cleavage. This cleavage...
  4. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...T7 promoter upstream of the left homology arm, perform in vitro transcription, and use the IVT product... and infundibulum. In vivo electroporation is performed immediately after injection. Image from Ohtsuka...contrast to standard microinjection, i-GONAD is performed directly on pregnant female mice using a small...
  5. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...repeating for each target. Alternatively, you can perform simultaneous detection with a mixture of primary...steps, in order to ensure your antibodies are performing their best and that the sample is not being degraded... P., & Yong, W. H. (2019). An Introduction to Performing Immunofluorescence Staining. In W. H. Yong (Ed...
  6. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...When performing restriction digests on plasmids at Addgene, we sometimes observe something odd in our...Running undigested DNA on a gel as a control when performing a diagnostic digest is a simple but effective... sequencing Addgene uses Illumina MiSeq NGS to perform whole plasmid sequencing of our plasmid samples...
  7. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...can be combined with CRISPR/Cas9 to efficiently perform genetic manipulations in animal models or cells...activity. In summary, the piggyBac™ transposon is a powerful technology that may be used for a variety of research...piggyBac™ for gene editing allows you to efficiently perform seamless genetic manipulations, while avoiding ...
  8. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...Read on for an in-depth breakdown of how to do perform restriction digests.   Overview of the restriction... common chemically-competent cells. You should perform, at minimum, two transformations after a ligation... 101 Posts Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse...
  9. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...what I think are the main tips for each step of performing and analyzing a gene edit using Addgene’s lentiviral...lentiCas9-blast. Anti-Cas9 Western blotting was performed on whole-cell extracts from stable pools of A549...blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell extracts from the initial stable...
  10. Finding Your Science Policy Path

    Type
    Blog Post
    ...and grow along the way. That’s what makes it so wonderful. The most challenging part of jumping into this...me, the Veterans Health Administration was the perfect position because my science policy ‘why’ was to...this question may change over time and that’s perfectly normal!  Action Items:  Start a career journal...
  11. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...phenotypes. These approaches may become even more powerful following a report that one can use linear repair...non-linked loci; pha-1(e2123) mutant worms are perfectly viable at 15 ºC, yet exhibit complete embryonic...recombination repair pathway. Reagents required to perform pha-1 co-conversion are available through Addgene...
  12. Pipetting Protocol

    Type
    Protocol
    ...the Volume on the Pipette Although each pipette performs the same function, the numbers are read differently...
  13. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...homologous recombination has not occurred. This powerful negative selection system ensures high editing...Transcriptional repression (CRISPRi) in bacteria As RNA interference does not function in bacteria, most efforts ...inMycobacterium tuberculosisusing an efficient CRISPR interference system. Nucleic Acids Res 44:e143–e143 . https...
  14. Pouring LB Agar Plates

    Type
    Protocol
    ...technique if you are working with any weird and wonderful organisms. While your samples are sterilizing ...
  15. Find and Share AAV Data with Addgene's New AAV Data Hub

    Type
    Blog Post
    ...long the scientist waited after injection before performing the experiment. Every entry has an accompanying... so that you can see for yourself how the AAV performed in that experimental setup. For example, this ...
  16. AAV ddPCR Titration

    Type
    Protocol
    ...Follow this protocol to perform digital droplet PCR (ddPCR) for purified AAV titration....
Showing: 221 - 240 of 631 results