Two-step "hot" PCR amplification of GC-rich avian c-myc sequences.
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 (Tm between 70 degrees and 74 degrees C), a two-step cycle that employs a 94 degrees C denaturation step and an annealing-elongation step between 70 degrees and 80 degrees C with or without formamide.
Schuchard
M
M
Dept. of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, MN 55905.
Sarkar
G
G
Ruesink
T
T
Spelsberg
T C
TC
D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D002645:Chickens; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D005260:Female; D005784:Gene Amplification; D016259:Genes, myc; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
D015342:DNA Probes; D004247:DNA