Sensitive and fast mutation detection by solid phase chemical cleavage

Hum Mutat. 1996;7(3):256-63. doi: 10.1002/(SICI)1098-1004(1996)7:3<256::AID-HUMU10>3.0.CO;2-1.

Abstract

We have developed a solid phase chemical cleavage method (SpCCM) for screening large DNA fragments for mutations. All reactions can be carried out in microtiterwells from the first amplification of the patient (or test) DNA through the search for mutations. The reaction time is significantly reduced compared to the conventional chemical cleavage method (CCM), and even by using a uniformly labelled probe, the exact position and nature of the mutation can be revealed. The SpCCM is suitable for automatization using a workstation to carry out the reactions and a fluorescent detection-based DNA sequencing system to analyze the cleaved fragments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins
  • Base Sequence
  • Biotin
  • DNA / genetics*
  • DNA Probes
  • Female
  • Genetic Techniques*
  • Humans
  • Molecular Sequence Data
  • Mutation*
  • Osmium Tetroxide
  • Polymerase Chain Reaction
  • Pyruvate Dehydrogenase Complex Deficiency Disease / genetics*
  • Sensitivity and Specificity
  • Sequence Deletion
  • Streptavidin

Substances

  • Bacterial Proteins
  • DNA Probes
  • Biotin
  • DNA
  • Streptavidin
  • Osmium Tetroxide