Experience on Sequencing of Mitochondrial DNA from 1200 Year Old Bone Using Cloning

Ancient bones have undergone natural decomposition and have been exposed to external environment for long period. Ancient DNA from old bone is usually fragmented. In addition, various kinds of inhibitors are co-extracted. All these may inhibit proper sequencing reaction. Cloning is regarded as the standard method when sequencing aDNA. When cloning, each clone from the same sample may not be of same sequence, and to exact consensus sequence may be difficult. Here we present our experience on 1200 year old bone from Russia, Primorsky Kray area. We have tried to sequence for HV I, II region of mtDNA using modified mini-primer set, which consisted of 7 set to cover the HV I, II. We cloned the PCR product and sequenced all the clones. Amplification efficiency and subsequent success rates were different for each mini primer set. Loci of variation that differ from consensus sequences were rather frequent, and the pattern were variable depending on sample. Except major polymorphic sites that are important when haplogroup designation, 16129 was the most frequent site that was discarded when extracting haplogroup designation.

발해 고인골의 mtDNA 자료입니다. 이 고인골은 발해 고분으로 추정되는 연해주 옥탸브리스키 군 체르냐찌노 5고분군의 고인골 14개를 분석한 것입니다. 연대는 대략 1200년~1400년 전으로 추정된다고 합니다. 제가 mtDNA 데이터는 데이터베이스를 마련하지 못해 별도의 분석은 하지 못하였으나 발해 고인골의 유전정보는 희귀한 자료이므로 소개해봅니다. 이 자료의 하플로그룹은 SNP를 검사하여 결정한 것은 아니며 HVR 데이터로 추정한 하플로그룹입니다. 

Open Access http://www.kjlm.or.kr/v1/bbs/board.php?bo_table=abstract&wr_id=114&no=1

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