Multiplex PCR for Three STR Loci of [GATA]n Repetition Unit and Its Forensic Application.

Korean J Leg Med. 1997 May;21(1):32-40. Korean.

Department of Forensic Medicine, Seoul National University College of Medicine

Multiplex PCR for three STRs of same repetition unit [GATA]n, 4804LR[D12S66], 27H39LR[DYS19] and 4815LR[D12S67] loci, was constructed for forensic application DNA was extracted from 200 unrelated Koreans and amplified with a mixture of polyacrylamide gel electrophoresis, so called Amp-FLP procedure. Three loci could be co-amplified in a reaction with easy, and reaction condition was not so quite different from that of each locus. The PCR products of each locus could be separated bp, and 4815LR from 241 bp to 281 bp, so these alleles of each locus could be separated on a single electrophoresis. A total of six alleles was noted in 4804LR and heterozygosity was 0.5764. The allele 11 and allele 12 were frequently noted with the frequency of 0.6225 and 0.1775, respectively. Sequencing was done for 2 alleles, and the exact size of the alleles and the repetition unit were confirmed. Through statistical analysis forensic applicability of the STR 4804LR locus was confirmed. For 4815LR and heterozygosity was 0.5764. The allele 11 and llele 12 were frequently noted with the frequency of 0.6225 and 0.1775, respectively. Sequencing was done for 2 alleles, and the exact size of the alleles and the repetition unit were confirmed. Through statistical analysis forensic applicability of the STR 4804LR locus was confirmed. For 4815LR locus the amplification was successful, but the separation of the alleles on routine polyacrylamide gel was not successful. Some alleles was hardly separable, some alleles did not match the allelic ladder exactly, so the interallele was suspicious. On sequencing gel the electrophoresis pattern was quite different with that of routine polyacrylamide gel. A total of 11 allele was noted in 4815LR and heterozygosity was 0.765. For the routine use of the 4815LR locus, more meticulous method for the separation of the alleles such as using automatic DNA sequencer was necessary.

Key words: multiple PCR, [GATA]n, STR, 4804LR, 27H39, 4815LR