1996 Volume 1 Issue 1 Pages 33-37
PCR amplified products on TH01 locus from bloodstain samples were analyzed by denatured polyacrylamide gel and were detected by silver staining. To adjust the PCR amplification condition on TH01 locus various annealing temperatures were tested and the optimum condition was revealed as 64°C or 66°C. Under the adopted typing procedure, the separation between allele 10 and allele 10-1, which is one base shorter than allele 10, were successfully performed.
The distribution of allele frequency on TH01 locus was examined of 501 unrelated Japanese. Seven alleles were observed ranging from allele 5 to 10 including allele 10-1 with allele 9 as the most common allele. No significant deviation from Hardy-Weinberg equilibrium was observed. Observed heterozygosity was 0.723 and expected heterozygosity was 0.712.
This system was applied to aged bloodstain samples and positive result was obtained from 5 out of 6 bloodstain samples left at room temperature for 16 years whose extracted DNA was partially decomposed.
