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 extra cted with organic solvent and purified by filtration. The filtrate sub sequently provides the mtDNA template for the PCR. The two hypervariab le segments of the mtDNA control region are amplified in four separate reactions. After a purification step to remove unincorporated PCR pri mers, amplified products are quantitated by capillary electrophoresis and subjected to cycle sequencing. The products are separated and anal yzed on an automated DNA sequencer The mtDNA sequences from the hair s haft match the mtDNA sequences from blood samples taken from the same donor.