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2013年50 No.1
Strategy and methods for sequencing mitochondrial genome
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Key Words:mitochondrial genome, longPCR, primerwalking, nextgeneration sequencing
Abstract:
This paper reviews strategy and methods for sequencing mitochondrial genomes. Two traditional sequencing methods are introduced; the clone librarybased sequencing method from physically purified mtDNA, and the PCR ampliconbased sequencing method, which includes primerwalking sequencing from long PCR amplicons and primerwalking sequencing from total DNA. For the application of nextgeneration sequencing strategies, there are total DNAbased methods, which include the parallel tagged sequencing (PTS) method based on preamplifying mtDNA and total DNA, and the transcriptomic sequencing method based on total RNA. From a practical perspective, which method is chosen depends on the scale of the research, size and preservative status of the sample, and financial support. The primer walking sequencing method based on longPCR amplicons is particularly suitable for small scale research on insect mitochondrial genomes. For large scale mitochondrial genome research, NGS technology is the best choice for saving time and effort.