REPLACR Mutagenesis: Replacing In Vitro Recombination Methods
Type
Blog Post
Published
Feb. 10, 2016, 3:30 p.m.
...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...