오래된 유해의 mtDNA 염기서열 분석률 향상에 관한 연구
Improvement of mtDNA Sequencing Analysis Rate from the Old Skeletal Remains
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초록
Mitochondrial DNA (mtDNA) sequencing is widely used in the identification of old skeletal remains or severely damaged biological samples. Recently, many studies on mtDNA sequencing using multiplex polymerase chain reaction (multiplex PCR) have been conducted as a means of improving the amplification efficiency by reducing the size of amplicons and the consumption of template DNA which is limited in quantity. This study aims to establish a less laborous and a more optimized method for mtDNA sequencing by using highly efficient and accurate multiplex PCR. Among Taq polymerases, TITANIUM Taq showed better results in success rate and reproducibility for multiplex PCR than i-Taq plus and AmpliTaq Gold. Using optimized multiplex PCR method with TITANIUM Taq and modified primer sets, it demonstrated a 95.0% success rate of mtDNA sequencing with 200 samples. This method shows tremendously increase success rate compared to the that of the 299 samples tested with the conventional method of S.M. Edson et. al. (63.7%). With this optimized multiplex PCR method, mtDNA sequencing with skeletal remains more than half of a century old can be successfully, and reproducibly analyzed.
Abstract
Mitochondrial DNA (mtDNA) sequencing is widely used in the identification of old skeletal remains or severely damaged biological samples. Recently, many studies on mtDNA sequencing using multiplex polymerase chain reaction (multiplex PCR) have been conducted as a means of improving the amplification efficiency by reducing the size of amplicons and the consumption of template DNA which is limited in quantity. This study aims to establish a less laborous and a more optimized method for mtDNA sequencing by using highly efficient and accurate multiplex PCR. Among Taq polymerases, TITANIUM Taq showed better results in success rate and reproducibility for multiplex PCR than i-Taq plus and AmpliTaq Gold. Using optimized multiplex PCR method with TITANIUM Taq and modified primer sets, it demonstrated a 95.0% success rate of mtDNA sequencing with 200 samples. This method shows tremendously increase success rate compared to the that of the 299 samples tested with the conventional method of S.M. Edson et. al. (63.7%). With this optimized multiplex PCR method, mtDNA sequencing with skeletal remains more than half of a century old can be successfully, and reproducibly analyzed.
참고문헌 (10)
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