PCR SD mutagenesis

PCR SD mutagenesis - Denature mix the products and anneale...

Info iconThis preview shows page 1. Sign up to view the full content.

View Full Document Right Arrow Icon
Site specific nucleotide substitution mutagenesis by PCR A B C A B C A B C You wish to change a single base in the above sequence at the site indicated by the arrow. A B C To do this you synthesize four primers, illustrated below. Two are identical to the sequence and correspond to the two ends of the fragment (the “outside” primers). The other two overlap the location that you want to mutagenize and include the spe- cific mutation (the “mutagenesis” primers, with the mutation indicated by a “blip” in the lines).
Background image of page 1
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: Denature, mix the products and anneale. One of the annealing products is shown below. This is the only one that would function as a template for PCR with only the outside primers. A B C Fill in with DNA polymerase PCR Set up two PCR reactions with the primers indicated below. They will produce the indicated products that will include the desired mutation. Add outside primers and PCR to amplify the mutant product A B C A B C A B C PCR C. S. Gasser, U. C. Davis, 2008...
View Full Document

This note was uploaded on 04/06/2011 for the course MCB 121 taught by Professor Gasser during the Winter '09 term at UC Davis.

Ask a homework question - tutors are online