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We narrowed to 130 results for: primer

Showing: 11 - 20 of 130 results

  1. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...mutagenic primer pair to generate an individual mutant, this technique uses mutagenic primers that tile...site and multiple primer sets tile across the protein or region of interest. A low primer-to-template ratio...How should I design my mutagenic primers? The key is to design primers that are the same sense as the strand...site-directed mutagenesis PCR that uses mutagenic primers to introduce all desired codon changes (step 1 ...1 in the image above). A second universal primer is used to synthesize a complementary mutant strand (... Synthesize mutant strand: A mix of degenerate primers and the high-fidelity Phusion polymerase are used...the newly synthesized DNA strand. Each degenerate primer set contains three consecutive randomized bases...

  2. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...shows guidelines for primer design; a desired insertion should be present in both primers. When deleting a...the same technique just using a large primer known as a megaprimer. Piecing together multiple PCR products...point mutation, you design two oligonucleotide primers with the desired mutation, then use those to PCR...discussed methods. As described above, you design two primers, each containing the desired mutations, to amplify.... In addition to containing the mutations, the primers also have some homology to each other; ~17 bases... a region, both primers should cover the newly joined ends surrounding the deletion. REPLACR-mutagenesis...nucleotide insertion; beyond this insertion size, primer synthesis is more expensive than gBlock synthesis...

  3. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...designed many primers for Sanger sequencing at Addgene. By using Snapgene’s “Import primers from a list”...Snapgene for additional verification experiments: Primer and restriction digest design Some regions of plasmids...list” function, we can detect all of the primers we have designed that will anneal to the plasmid sequence...sequence. We can then quickly choose which primers we should use to further verify the difficult regions and...in our plasmid maps, including our most popular primers and certain restriction enzymes, but you will also...

  5. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...overlapping region of the primer to 40-60 bp. It is always a good idea to perform primer optimization, especially...large genome, you can recheck your primer design or optimize your primers or PCR conditions. Purifying DNA...complementary to standard sequencing primers. If not, design primers to sequence your insert. When you get...1-2 ug of the vector for digestion. PCR Design primers that overlap at least 24 bp with the template and...help you with that, they can be open source (i.e. primer3) or provided by the major companies selling PCR...

  6. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...Sequence” tabs. Primers This tab contains a table listing commonly used primers detected in a given...maps. Clearly Annotated Features, Enzymes, and Primers - The new maps have the identities of most features...functionalities. You can now hover over any feature, enzyme, primer, or ORF to learn more information about it. For...given nucleotide sequence. The table includes primer names, sequences, binding site locations, lengths, ..., and direction. Clicking on a particular primer here also highlights it in the “Map” and “Sequence” tabs...

  7. Synthetic Biology - Assembly Standards Guide

    Type
    Collection
    ...sequences for each (these can be built into PCR primers when creating a new part, with an additional 5'...suffix will need to be reverse complimented for primer design. The restriction enzymes used for assembly...

  8. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...fragments can be created via PCR amplification with primers that contain the appropriate homologous sequences.... NEB recommends an overlap of 15-40 bp, with a primer melting temperature greater than 48℃. Both Snapgene...Snapgene and NEB have tools that help you design primers for PCR amplification of fragments to incorporate...demonstration of how to use Snapgene’s program to design primers for Gibson Assembly. For a simple example of using...scars as depicted below. First, you need to design primers to amplify the two fragments while also including...of colonies. Excluding the time it takes to get primers, you can have your construct made in 5 days. One... between experiments. In Gibson assembly, long primers for each fragment must be designed and ordered,...

  9. 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 ...

  10. 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...
Showing: 11 - 20 of 130 results