HLA-DRB1 Study of DNA from Ancient Human Skeleton by Sequence-based Typing / 체질인류학회지

The analysis of ancient human DNA is increasingly used recently in the study of anthropology and human evolution. Although mitochondrial DNA and Y chromosomal DNA has commonly been the target in the field of human DNA study, HLA analysis of ancient human DNA is extremely rare. This study aimed to develop the PCR method of ancient human DNA for analyzing the sequence of HLA. Authors established a new method for HLA-DRB1 analysis by sequence-based typing. Alleles of HLA-DRB1 were analyzed and typed by sequencing with DNA of ancient human skeletons from Korea and Mongolia 3000-500 years ago. The types of HLA-DRB1 were determined by comparing the sequences with those of HLA database (http://www. ebi.ac.uk/Tools/blast2/nucleotide.html). The alleles of HLA-DRB1 of ancient human DNA from Korea and Mongolia were classified by types. The frequencies of HLA-DRB1 types of Mongolia were also presented according to the geography such as West, Central, East, and North. In summary, our method was successful in the analyzing the type of HLA-DRB1 from DNA of ancient human bones. Authors anticipate that many researchers could do their research in a better way to get the genetic information for the kinship analysis between individuals or communities from ancient human bones.

Establishment of PCR Reaction Condition for Highly Successful Ancient DNA PCR / 체질인류학회지

The ancient bone DNA analysis essentially requires PCR amplification of the targeting genes of study due to the limitation of the ancient bone sample and DNA amounts. In contrast to the fresh living human DNA, it is common to face failing in amplifying the poorly preserved ancient DNA after death. Therefore, the optimized PCR methods appropriate for ancient DNA are required. However, there is no report to date that a systemic investigation of enhanced PCR amplification methods suitable for ancient samples has been conducted Approximately 500~3,300-year-old Korean and Mongolian ancient bones that are resistant to PCR were selected and an extensive number of PCR conditions were systematically investigated for the comparison of PCR success rates. For the PCR analysis, a mitochondrial DNA fragment as a multicopy DNA and a M175 Y chromosome biallelic marker DNA fragment as a single copy DNA that is the marker of the prevalent Y haplogroup (haplogroup O) in Korea were targeted. The identity of the amplified products were confirmed by DNA sequencing. Through this study, we established the optimized PCR conditions for the highly successful amplification of ancient bone DNAs. This estabilished method allowed for the successful amplification of mitochondrial DNAs from all the ancient bone samples tested and the amplification by 50% success rates in the amplification of M175 Y chromosome biallelic marker DNA but with the highest success rates. These results demonstrate that the optimized PCR condition will be useful for the promising ancient DNA analysis in the fields of molecular genetic anthropological studies.