Abstract
A D1S80 typing method using 310 Genetic Analyzer is described. To improve the resolution in capillary electrophoresis, we designed new primers, which amplicon lengths were 125 bp smaller than those obtained with Kasai's original primers. The PCR amplification was performed using a DNA polymerase for long and accurate (LA) PCR to enhance the amplification of the larger allele of a heterozygote sample. When large amounts of DNAs were amplified, we observed “stutter-like” peaks that may be derived from the incomplete denaturation of the PCR products before the injection into the 310 Genetic Analyzer. However, “stutter-like” peaks were not detected, when the PCR products were diluted. The heterozygote peak height ratio (HPR), sensitivity, detection limit ratio in the mixed DNAs, sizing precision, and the concordance of the newly designed primers with Kasai's original primers were evaluated. Our D1S80 typing method was confirmed to be a reliable and useful method.
