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Showing: 21 - 30 of 99 results
  1. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...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...
  2. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...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...
  3. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...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...
  4. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...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...
  5. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...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...
  6. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...genome browser and Primer3 to design primers that amplified a 500-600 bp product. The primers I designed (listed...Dilution Lentivirus Production Websites Primer3: for Designing Primers UCSC Genome Browser: For eploring genomic...most difficult part of the assay was designing primers flanking my edits in genomic DNA and doing PCR ...samples (data not shown). Pro-Tips: 1.Designing primers against genomic DNA to amplify the gRNA targets...bands using the conditions above. 2. Design your primers so that the digestion site isn’t in the middle ...
  7. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...applications, results can be affected by DNA quality and primer design, particularly for amplicons under 100 bp...technique, courtesy of CAR15.) Each droplet contains a primer and probe set for both the gene of interest and... Chain Reaction (PCR) Protocol: How to Design a primer  Molecular Biology Reference Molecular Cloning ...
  8. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...are titered by droplet digital PCR (ddPCR) using primers and probes targeting the ITR elements and an internal...incubated for 72 hours, and genomic DNA is extracted. Primers and probes targeting integrated copies of the lentiviral...the final viral preparation undergoes PCR with primers targeting the transfer plasmid used in the transfection...
  9. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...selectable markers for use in other cell types. Primer Binding Site A short single-stranded DNA sequence...initiation point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids...
  10. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    Published
    Nov. 21, 2024, 10:47 p.m.
    ...Electrophoresis Bacterial Transformation How to Design a Primer Creating Bacterial Glycerol Stocks Inoculating ... Videos What is a Plasmid? How to Design Primers for PCR Agarose Gel Electrophoresis Inoculating...Analysis We also introduced guides — practical primers to help users get started with tools like CRISPR...
Showing: 21 - 30 of 99 results