Quantitative analysis of mitochondrial DNA deletion in paraffin embedded muscle tissues from patients with KSS and CPEO

Biochim Biophys Acta. 1997 May 24;1360(3):193-5. doi: 10.1016/s0925-4439(97)00022-7.

Abstract

The percentage of common deletion of mitochondrial DNA (mtDNA) was determined quantitatively by a PCR-based, non-radioactive method in DNA extracted from formalin-fixed, paraffin-embedded skeletal muscle tissues from two patients with Kearns Sayre syndrome (KSS) and one with chronic progressive external ophthalmoplegia (CPEO). The method involved PCR cycle titration of wild-type and deleted mtDNA in parallel, staining of gel bands with the sensitive fluorescence dye SYBR Green I, and quantitation of intensity on a computer screen by the NIH image program. We determined 75% and 71% common deletion of mtDNA in the KSS patients and 35% in the CPEO patient.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • DNA, Mitochondrial / analysis*
  • Female
  • Humans
  • Kearns-Sayre Syndrome / genetics*
  • Male
  • Mitochondria, Muscle / metabolism*
  • Mutation
  • Ophthalmoplegia, Chronic Progressive External / genetics*
  • Paraffin Embedding
  • Polymerase Chain Reaction
  • Sequence Deletion

Substances

  • DNA, Mitochondrial