The kinship was analyzed genetically on the three 2000 year old ancient human bones and teeth excavated in Mongolia. The samples were processed in a clean room to prevent the contamination from modern human DNA. The DNA extraction and purification was done with ion-exchange column kit (Qiagen G-tip 20G, USA). The PCR was done with purified DNAs from ancient human bones for paternal Y-SNP haplogroup, maternal mtDNA haplogroup, and autosomal short tandem repeats (STR). Two samples belonged to the maternal D major haplogroup, which is one of the most frequent types in the present North East Asia. One of them, showing male genotype, belonged to the paternal C major haplogroup, which is also one of the most frequent types in the present North East Asia. The remaining one belonged to the paternal R major haplogroup, frequent in the present Europe, and the maternal U haplogroup, frequent in the present Europe and East Mediterranean. The repeated results were consistent in the autosomal STR PCR. The STR data were analyzed with DNA-VIEW program (http://www.dna-view.com), which showed no close kinship among the three ancient humans. Our method was successful in the analyzing kinship among ancient human bones, which has been possible in few restricted laboratories in the World. Authors anticipate that many researchers could do their research in a better way to get the genetic information from ancient human bones.
DNA ; DNA, Mitochondrial ; Environment, Controlled ; Europe ; Far East ; Genotype ; Humans ; Male ; Microsatellite Repeats ; Mongolia ; Polymerase Chain Reaction ; Tooth

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.
Alleles ; Anthropology ; DNA ; DNA, Mitochondrial ; Geography ; HLA-DRB1 Chains ; Humans ; Korea ; Mongolia ; Polymerase Chain Reaction ; Skeleton