Sequence Analysis of Mitochondrial DNA Hypervariable Regions Using Infrared Fluorescence Detection | Zendy

David L. Steffens | Zendy; Reena Roy | Zendy

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Sequence Analysis of Mitochondrial DNA Hypervariable Regions Using Infrared Fluorescence Detection

Author(s) -

David L. Steffens,

Reena Roy

Publication year - 1998

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/98246pf01

Subject(s) - hypervariable region , mitochondrial dna , primer (cosmetics) , biology , genetics , mtdna control region , dna , microbiology and biotechnology , polymerase chain reaction , coding region , sequence (biology) , dna sequencing , sequence analysis , genome , computational biology , gene , chemistry , genotype , organic chemistry , haplotype

The non-coding region of the mitochondrial genome provides an attractive target for human forensic identification studies. Two hypervariable (HV) regions, each approximately 250-350 bp in length, contain the majority of mitochondrial DNA (mtDNA) sequence variability among different individuals. Various approaches to determine mtDNA sequence were evaluated utilizing highly sensitive infrared (IR) fluorescence detection. HV regions were amplified either together or separately and cycle-sequenced using a Thermo Sequenase protocol. An M13 universal primer sequence tail covalently attached to the 5' terminus of an amplification primer facilitated electrophoretic analysis and direct sequencing of the amplification products using IR detection.

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