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Clin Biochem. 1998 Nov;31(8):627-32.

A protocol for detection of mitochondrial DNA deletions: characterization of a novel deletion.

Author information

  • 1Department of Pediatrics, University of British Columbia, Vancouver, Canada. marioncm@interchange.ubc.ca

Abstract

OBJECTIVES:

To develop a protocol capable of identifying deletions in mitochondrial DNA and use it to identify the breakpoints of a mtDNA deletion in a patient with chronic progressive external ophthalmoplegia (CPEO).

DESIGN AND METHODS:

Deletions in mtDNA were identified by a combination of long range PCR and Southern blotting. The precise breakpoints were determined by automated DNA sequencing.

RESULTS:

A series of DNA samples from patients with suspected mitochondrial disease was subjected to a protocol, which combines long range PCR and Southern blotting. We found a unique deletion in a patient with CPEO and we identified the precise location of this deletion through DNA sequencing.

CONCLUSIONS:

Long range PCR has the advantages of speed, minimal samples requirements, and sensitivity. Southern blotting is better able to evaluate heteroplasmy and detect duplications. We suggest a protocol that enables us to identify precisely the breakpoints in a unique mutation of mtDNA in a patient with CPEO.

PMID:
9876894
[PubMed - indexed for MEDLINE]
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