The feasibility of PCR-based diagnosis of Prader-Willi and Angelman syndromes using restriction analysis after bisulfite modification of genomic DNA

Mol Genet Metab. 2000 Jan;69(1):81-3. doi: 10.1006/mgme.1999.2941.

Abstract

We have developed a novel PCR-based method for studying DNA methylation in the proximal region of 15q, using restriction analysis after bisulfite treatment of genomic DNA. This protocol can be used for the diagnosis of Prader-Willi and Angelman syndromes. Unlike the recently reported methylation-specific PCR protocol, our method avoids the use of multiplex amplification, thus overcoming the need to adjust relative primer amounts and the risk of obtaining false-negative results.

MeSH terms

  • Angelman Syndrome / diagnosis*
  • Angelman Syndrome / genetics
  • Autoantigens / genetics
  • Chromosomes, Human, Pair 15 / genetics
  • CpG Islands / genetics
  • DNA Methylation*
  • DNA Primers / genetics
  • Deoxyribonucleases, Type II Site-Specific / metabolism*
  • False Negative Reactions
  • Genetic Testing / methods
  • Genome
  • Genomic Imprinting / genetics
  • Humans
  • Polymerase Chain Reaction / methods*
  • Prader-Willi Syndrome / diagnosis*
  • Prader-Willi Syndrome / genetics
  • Ribonucleoproteins, Small Nuclear*
  • Sulfites / metabolism*
  • snRNP Core Proteins

Substances

  • Autoantigens
  • DNA Primers
  • Ribonucleoproteins, Small Nuclear
  • Sulfites
  • snRNP Core Proteins
  • Deoxyribonucleases, Type II Site-Specific
  • GCGC-specific type II deoxyribonucleases