title: Two-step "hot" PCR amplification of GC-rich avian c-myc sequences.
abstract: 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.
authors.affiliation: Dept. of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, MN 55905.
authors.forename: M
authors.initials: M
authors.lastname: Schuchard
authors.forename: G
authors.initials: G
authors.lastname: Sarkar
authors.forename: T
authors.initials: T
authors.lastname: Ruesink
authors.forename: T C
authors.initials: TC
authors.lastname: Spelsberg
mesh_terms: 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*
chemical_list: D015342:DNA Probes; D004247:DNA