Quantitative analysis of mitochondrial DNA deletion in paraffin embedded muscle tissues from patients with KSS and CPEO.
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.
Kim
S H
SH
Department of Biology, Taegu University, Kyungbuk Kyungsan-Si, South Korea. SangKim@biho.taegu.ac.kr
Chi
J G
JG
Reith
A
A
Kadenbach
B
B
D000328:Adult; D004272:DNA, Mitochondrial / Q000032:analysis*; D005260:Female; D006801:Humans; D007625:Kearns-Sayre Syndrome / Q000235:genetics*; D008297:Male; D008931:Mitochondria, Muscle / Q000378:metabolism*; D009154:Mutation; D017246:Ophthalmoplegia, Chronic Progressive External / Q000235:genetics*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion
D004272:DNA, Mitochondrial