Competitive polymerase chain reaction for the quantification of N-myc gene copy number in neuroblastoma

Tumour Biol. 1996;17(5):262-70. doi: 10.1159/000217988.

Abstract

An absolute quantification method for the N-myc gene copy number of neuroblastoma specimens was established by applying the competitive polymerase chain reaction (cPCR). The competitor plasmid (pZH2) lacking an MluI site in the exon 2 was constructed to distinguish two product species amplified from genomic DNA and the competitor plasmid. By using this cPCR system, we could obtain qualitative results within 1 day, i.e. amplified or unamplified, and quantitative results by using radiolabelled nucleotides within 4 days. The copy numbers of N-myc in 47 neuroblastoma specimens by cPCR correlated well with those by Southern hybridization (r = 0.85). We conclude that cPCR is a simple and rapid method, requires only a small amount (200 ng) of sample DNA, and is expected to be used for prognostic evaluation in neuroblastomas.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Breast Neoplasms / genetics
  • Gene Dosage*
  • Genes, myc*
  • Humans
  • Molecular Sequence Data
  • Neuroblastoma / genetics*
  • Polymerase Chain Reaction*

Associated data

  • GENBANK/X00351
  • GENBANK/Y00664