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  1. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...technique is frequently called allelic exchange. Here, a mutated allele introduced from a conjugative ... an allelic exchange experiment? Find the pTOX plasmids here! Continued development of new allelic exchange...off-target mutations. Ultimately, successful allelic exchange requires two sequential homologous recombination...counterselection vector is the pDS132 sacB-based allelic exchange vector. This vector contains sacB, an chloramphenicol...resistant colonies isolated are those where the allelic exchange vector has undergone single crossover and ...develop these negative selection markers for allelic exchange. To do this, we performed the following modifications...exchange vectors Allelic exchange is a great technique for the scarless modification of bacterial genomes...
  2. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...specific purposes.     Chromoproteins for allelic exchange Figure 3: pTOX plasmids expressing amilCP...chromoproteins to create negative selection markers for allelic exchange. These vectors contain a toxin gene (regulated...rhamnose-inducible promoter) and a chromoprotein gene. Allelic exchange occurs through two sequential homologous recombination...encode the tsPurple chromoprotein.  Find the allelic exchange vectors here! ShadowR, a novel FLIM-FRET acceptor...M., & Waldor, M. K. (2019). A New Suite of Allelic-Exchange Vectors for the Scarless Modification of Proteobacterial...
  3. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...stranded breaks, in which nucleotide sequences are exchanged between two similar or identical molecules of ...Figure 4: A targeting vector to create a floxed allele for a conditional knockout. This experimental design...include loxP sites to create conditional floxed alleles. Although CRISPR is very good at making knockouts...
  4. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ... could recapitulate or rescue known pathogenic alleles, or introduce a premature stop codon to render ... their RESCUE (RNA Editing for Specific C-to-U Exchange) system (Abudayyeh et al., 2019). To avoid some...
  5. Antibody Production

    Type
    Collection
    ...affinity chromatography, antibodies undergo a buffer exchange using centrifugal columns. The final formulation...Whenever possible, the experiment is performed in parallel with a previous lot of recombinant antibody or...
  6. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...activation of the Gα subunit. The activated Gα exchanges bound GDP with GTP, resulting in the disassociation...created an open source resource called PRESTO-TANGO (parallel receptorome expression and screening via transcriptional...
  7. Guide to Using Pooled Libraries

    Type
    Guide
    ...consult these links: NGS Company Resources Science Exchange Scientist (formally Assay Depot) Resources Find...Sequencing (NGS) on a control sample (no selection) In parallel, apply library Apply selection; most cells will...Sequencing Next-generation sequencing allows for parallel sequencing of millions of DNA fragments simultaneously...
  8. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...sites, and provides a modular system for easier exchange and re-use of existing elements. DNA sequences...expression. The capability to turn RUSH and CRUSH alleles off or on enables this method to rapidly address...carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown in eyes and clonogenic...mammalian cells. These vectors were designed for parallel testing and screening of constructs in multiple...
Showing: 1 - 9 of 9 results