Two-step 'hot' PCR amplification of GC-rich avian c-myc sequences

M. Schuchard, G. Sarkar, T. Ruesink, T. C. Spelsberg

Research output: Contribution to journalArticle

10 Scopus citations

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 (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 languageEnglish (US)
Pages (from-to)390-392+394
JournalBioTechniques
Volume14
Issue number3
StatePublished - Jan 1 1993

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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    Schuchard, M., Sarkar, G., Ruesink, T., & Spelsberg, T. C. (1993). Two-step 'hot' PCR amplification of GC-rich avian c-myc sequences. BioTechniques, 14(3), 390-392+394.