Heng Nicholas C K, Stanton Jo-Ann L
Faculty of Dentistry, Sir John Walsh Research Institute, University of Otago, Dunedin, New Zealand.
Methods Mol Biol. 2010;666:197-218. doi: 10.1007/978-1-60761-820-1_14.
For over 30 years, the chain termination method of DNA sequencing (commonly known as Sanger sequencing) has been the mainstay of any DNA sequencing project. In the past, whole-genome sequencing employing exclusively Sanger chemistry has been a labor-intensive and costly exercise and an option unfeasible for the average research group. However, within the last 4 years, the introduction of three high-throughput sequencing technologies (454, SOLiD, and Illumina) has revolutionized genomics by facilitating unprecedented levels (up to gigabasepairs) of reliable DNA sequence output in a relatively short time frame and at a much lower cost per sequenced basepair. Here, we provide laboratory and bioinformatic protocols that will allow the average research group to undertake high-throughput sequencing of oral bacterial genomes using the Roche Genome Sequencer FLX System which employs 454 pyrosequencing technology.
30多年来，DNA测序的链终止法（通常称为桑格测序）一直是任何DNA测序项目的支柱。过去，仅采用桑格化学法进行全基因组测序是一项劳动密集型且成本高昂的工作，对于普通研究团队来说是不可行的选择。然而，在过去4年里，三种高通量测序技术（454、SOLiD和Illumina）的引入彻底改变了基因组学，它们能够在相对较短的时间内以空前的水平（高达数十亿碱基对）产出可靠的DNA序列，且每个测序碱基对的成本要低得多。在此，我们提供了实验室及生物信息学方案，这将使普通研究团队能够使用采用454焦磷酸测序技术的罗氏基因组测序仪FLX系统对口腔细菌基因组进行高通量测序。
