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Rapid mitochondrial DNA typing using restriction enzyme digestion of polymerase chain reaction amplications followed by capillary electrophoresis separation with laser‐induced fluorescence detection
Author(s) -
Butler John M.,
Wilson Mark R.,
Reeder Dennis J.
Publication year - 1998
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150190120
Subject(s) - mitochondrial dna , restriction enzyme , polymerase chain reaction , capillary electrophoresis , microbiology and biotechnology , biology , restriction fragment length polymorphism , typing , hypervariable region , genetics , dna , gel electrophoresis , mtdna control region , restriction digest , population , gene , allele , demography , sociology , haplotype
The polymorphic control region of mitochondrial DNA (mtDNA) is becoming more commonly used in forensic applications to differentiate among individuals in a population. Two hypervariable regions (HV1 and HV2) are often sequenced following amplification of the mtDNA via the polymerase chain reaction (PCR). More rapid screening assays would reduce both the effort and the expenses of comparing two samples. A methodology has been developed that first uses restriction endonuclease digestion of the PCR‐amplified mtDNA using Rsa I and Mn /I and then capillary electrophoresis (CE) to separate and size the PCR‐RFLP fragments. This rapid procedure offers an alternative method for screening of polymorphisms in amplified mtDNA samples. In addition, the presence of a T→C transition at position 16189, which gives rise to the so‐called “C‐stretch” in HV1, may be predicted from the presence of nonspecific PCR products in the CE results.