Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts

Techniques have been developed for extracting, amplifying and directly sequencing mitochondrial DNA (mtDNA) from human hair shafts. The hair shaft is ground in a glass micro-tissue grinder, and the DNA is extracted with organic solvent and purified by filtration. The filtrate subsequently provides t...

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Bibliographic Details
Published in:	BioTechniques Vol. 18; no. 4; p. 662
Main Authors:	Wilson, M R, Polanskey, D, Butler, J, DiZinno, J A, Replogle, J, Budowle, B
Format:	Journal Article
Language:	English
Published:	England 01-04-1995
Subjects:	Base Sequence DNA Primers DNA, Mitochondrial - isolation & purification Fluorescent Dyes Hair - chemistry Humans Molecular Sequence Data Polymerase Chain Reaction Sensitivity and Specificity Sequence Analysis, DNA
Online Access:	Get more information
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Summary:	Techniques have been developed for extracting, amplifying and directly sequencing mitochondrial DNA (mtDNA) from human hair shafts. The hair shaft is ground in a glass micro-tissue grinder, and the DNA is extracted with organic solvent and purified by filtration. The filtrate subsequently provides the mtDNA template for the PCR. The two hypervariable segments of the mtDNA control region are amplified in four separate reactions. After a purification step to remove unincorporated PCR primers, amplified products are quantitated by capillary electrophoresis and subjected to cycle sequencing. The products are separated and analyzed on an automated DNA sequencer. The mtDNA sequences from the hair shaft match the mtDNA sequences from blood samples taken from the same donor.
ISSN:	0736-6205