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

M. R. Wilson, D. Polanskey, J. Butler, J. A. DiZinno, J. Replogle, B. Budowle

Research output: Contribution to journalArticle

171 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)662-664+666
JournalBioTechniques
Volume18
Issue number4
StatePublished - 1 Jan 1995

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Mitochondrial DNA
Hair
Amplification
Polymerase Chain Reaction
DNA sequences
Capillary electrophoresis
Grinding mills
DNA
Capillary Electrophoresis
Organic solvents
Purification
Glass
Blood
Tissue

Cite this

Wilson, M. R., Polanskey, D., Butler, J., DiZinno, J. A., Replogle, J., & Budowle, B. (1995). Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts. BioTechniques, 18(4), 662-664+666.
Wilson, M. R. ; Polanskey, D. ; Butler, J. ; DiZinno, J. A. ; Replogle, J. ; Budowle, B. / Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts. In: BioTechniques. 1995 ; Vol. 18, No. 4. pp. 662-664+666.
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Wilson, MR, Polanskey, D, Butler, J, DiZinno, JA, Replogle, J & Budowle, B 1995, 'Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts', BioTechniques, vol. 18, no. 4, pp. 662-664+666.

Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts. / Wilson, M. R.; Polanskey, D.; Butler, J.; DiZinno, J. A.; Replogle, J.; Budowle, B.

In: BioTechniques, Vol. 18, No. 4, 01.01.1995, p. 662-664+666.

Research output: Contribution to journalArticle

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AB - 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.

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Wilson MR, Polanskey D, Butler J, DiZinno JA, Replogle J, Budowle B. Extraction, PCR amplification and sequencing of mitochondrial DNA from human hair shafts. BioTechniques. 1995 Jan 1;18(4):662-664+666.