Analysis of Mixed Samples using Automatic Sequencer and Establishment of Quantitative PCR.

Korean J Leg Med. 1997 May;21(1):23-31. Korean.

Department of Forensic Medicine and Internal Medicine, Seoul National University College of Medicine

To know the amplification pattern according to relative concentration ratio in mixed samples, two STRloci, vwF locus and MBP locus and two VNTR loci, D1S80 locus and d17S5 locus were amplified in DNA with various concentration of two individuals were easily identified. But when the concentration of one person were lowered to 1/20-1/40 of the other's the intensity of product bands diminshed and hardly discernible. Also different amplification efficiency according to the template length was noted, especially in VNTR loci. Using automatic sequencer and RFLP scan program, the intensity OD of each PCR product band could be calculated, and this correlates the felative amplification efficiency of each allele. By using this we could construct quantitative PCR for the mixed samples. This could be used in practical case work for forensic purpose, and also be a valuable candidate for 'chimerism detection' in case of bone marrow transplatation.

Key words: Mixed sample. Amplification efficiency, automatic sequencer, quantitative PCR, bone marrow transplantation