Sequence analysis for HV I region of mitochondrial DNA using WGA (Whole Genome Amplification) method.

The hypervariable (HV) region in mitochondrial DNA (mtDNA) has been studied for forensic identification, DNA analysis of mother-child relationship and comparison between ethnic groups. Meanwhile, the WGA (Whole Genome Amplification) method is a technique to amplify genome DNA, and it may be useful for analyzing degraded or limited DNA. However, we need to make a comparative study to confirm whether or not amplified DNAs have the same sequence as the original DNA because the amplification procedure may have some influence on sequencing. We sequenced mtDNA HV I region using amplified human DNAs by WGA method and original human DNAs, and compared the results. DNAs were extracted from four tissue samples from each of 10 healthy volunteers: blood, oral mucosa, nail and hair shaft. WGA was then performed and sequences were analyzed using the amplified samples. After sequencing of the PCR products, the results were compared with the Anderson reference sequence. For each individual, DNA sequences of mtDNA HV I region from the four tissues were the same and WGA amplified samples also showed identical results. Although the WGA procedure may have little influence on the data, we still have to investigate the effectiveness and technical limitation of the method.