NCJ Number
181273
Journal
Legal Medicine Volume: 1 Issue: 2 Dated: April 1999 Pages: 61-67
Date Published
1999
Length
7 pages
Annotation
Because the typing of nuclear DNA from hair shafts has often been unsuccessful, the authors tried to type one of the nuclear DNA loci, HLA-DQA1, from hair shafts using an efficient cetyl-trimethyl ammonium bromide (CTAB) precipitation for DNA purification and a sensitive semi-nested PCR.
Abstract
After thorough washing with ethanol and water, hair shafts were digested by proteinase K in the presence of dithiothreitol, followed by a purification step including CTAB-DNA precipitation. The specific region of HLA-DQA1 gene was amplified by the semi-nested PCR, and amplified products were cloned and sequenced. The HLA-DQA1 genotype was determined by comparing the sequence to the known sequence of each allele. All genotypes of HLA-DQA1 were successfully typed with hair shafts from six known heterozygotes, although one of them showed the predominant appearance of one allele. For correct typing, a template DNA equivalent to a hair shaft of 5 or 10 cm in length was necessary. Without the CTAB-DNA precipitation step, DNA extra from hair shafts inevitably contained enough melanin to inhibit PCR. The authors conclude that hair shafts can be used for the typing of nuclear DNA loci. 23 references, 1 table, and 4 figures