A new two-step cycle PCR method has been developed for amplification of GC-rich DNA sequences. Using this method, termed 'hot PCR', 111 and 179 bp regions (GC contents of 74% and 76%, respectively) of the avian c-myc proto-oncogene were specifically amplified from cloned and genomic DNA. This method uses high-melting primers (T(m) between 70° and 74°C), a two-step cycle that employs a 94°C denaturation step and an annealing-elongation step between 70° and 80°C with or without formamide.
|Original language||English (US)|
|State||Published - 1993|
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)