Computer Programs for PCR Primer Design and Analysis

Heat will separate or &quote;melt&quote; double-stranded DNA into single-stranded DNA by disrupting its hydrogen bonds. T _m (melting temperature) is the temperature at which half the DNA strands are single-stranded and half are double-stranded. T _m characterizes the stability of the DNA hybrid formed between an oligonucleotide and its complementary strand and therefore is a core parameter in primer design. It is affected by primer length, primer sequence, salt concentration, primer concentration, and the presence of denaturants (such as formamide or DMSO).