Designing PCR Primers Painlessly

Abstract

Students design primers using web-based tools along with a printed sequence of a prokaryotic gene showing both DNA strands and translation of the coding strand into amino acids. The melting temperature of each primer is determined and the pair closest in temperature can be used to amplify and clone a gene into a vector for a one-semester laboratory exercise. By working through this primer design exercise, students see first hand what is involved in the process of amplifying DNA.