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We narrowed to 129 results for: primer

Showing: 21 - 30 of 129 results
  1. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...design guide RNAs, donor templates and sequencing primers for quality control. At such a scale, automation... project was spent selecting guides, designing primers, and running analyses. New team members have taken...CrispyCrunch, or you can input pre-existing guides for primer design. The ecosystem of CRISPR tools Before deciding...tools we found performed batch guide design, batch primer design or batch analysis. We wanted to relieve ...lines of code, we enabled it to return guides and primers in batch, optimized for HDR. Compared to wealth...inputting the target regions Design donor RNA Design primers Review and order reagents Perform your wet lab ...it uses standard tools and databases, including primer3, biopython, bwa, samtools, bowtie, UCSC genome ...
  2. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...universal primers can be used to titer any lentiviral prep that shares that specific feature. Primers targeting...converted to cDNA and then quantified using qPCR primers targeting specific viral components such as LTRs...transgene itself. Many researchers prefer to design primers that target the common features of viral vector...dilutions of a viral prep, genomic DNA isolated, and primers targetting viral components or the transgene itself...
  3. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ...and the reverse primer is located in the 5’ end of the multi-cloning site. The primers used to amplify...and vector are first PCR amplified. The forward primer used to amplify the vector is located in the 3’...should perform colony PCR with vector specific primers to confirm that the products are the correct size...
  4. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...amplified by a particular primer pair. Then, by leveraging qPCR data from multiple primer pairs, one can infer...build new registers. The primer binding sites can also be used to design primers to read states by qPCR ... each state of a register enables the design of primer pairs that PCR amplify in some states but not others... regular PCR may not be very informative – each primer pair will amplify even if its target states are... promoters, and genes) were built with flanking primer binding sites (designed to have 20-30 nt length...
  5. Pushing the Limits of DNA Assembly

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    Blog Post
    ...homology can be introduced to fragments via PCR using primers that contain the regions of homology. A powerful...interface can guide you through both experimental and primer design based on fragment sequences and the polymerase... enzymes are adjusted. Appropriate fragment and primer design results in covalently joined, scarless DNA...search Addgene plasmids by gene here), designing primers with your desired mutation, performing PCR to generate...
  6. Design Tips for Prime Editing

    Type
    Blog Post
    ...of the primer binding site, starting with a length of about 13 nt (Anzalone et al., 2019). Primer binding...consists of a CRISPR gRNA extended to include a primer binding site (PBS) complementary to the genome ... 2: Starting lengths for optimizing your pegRNA primer binding site (PBS) and reverse transcriptase template...minimize unwanted intra-RNA base pairing with the primer binding site (Nelson et al., 2022). PE7: This editor...
  7. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...for binding sites between the primers and the AAV genome repeats. Primer annealing can be affected by... 61° C improved assay reliability. Different primers can have different annealing efficiencies (Wang... et al., 2013). Although you can optimize your primers for each individual sample, this will reduce convenience...each sample with be quantified with a different primer pair. Ct values vary based on the amount of starting...
  8. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...Remember the primers you designed to generate your pCAG-EGXXFP plasmid? They are the perfect primer sets to...and has the bonus benefit of supplying you with primers for validating your edit in mice once they have...vitro screening process, you should first develop primers to the approximately 500 base pair sequence surrounding...to genotype your pups. A simple PCR with these primers will let you know if there are any major deletions...
  9. Antibody Production

    Type
    Collection
    ...residual plasmid is extracted and amplified with primers targeting the heavy chain and/or light chain variable...
  10. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...extracted and subject to isothermal amplification by primers that target the E and the N2 gene of SARS-CoV-2...reverse-transcribed target sequence that are present close to the primer recognition sites and are added separately. Using...fluorescence upon cleavage. Upon amplification by the RPA primers, the Cas12a-crRNA binds to the complementary region...sequence is amplified by PCR/RPA using biotinylated primers. These are then immobilized with streptavidin coated...non-PAM regions, PAM can be engineered into the primers. This method does not depend on collateral cleavage...
Showing: 21 - 30 of 129 results