Department of Ecology and Evolutionary Biology, University of Toronto, 25 Willcocks St,, Toronto, Ontario, M5S 3B2, Canada.
Gigascience. 2012 Jul 12;1(1):8. doi: 10.1186/2047-217X-1-8.
The recent rise in speed and efficiency of new sequencing technologies have facilitated high-throughput sequencing, assembly and analyses of genomes, advancing ongoing efforts to analyze genetic sequences across major vertebrate groups. Standardized procedures in acquiring high quality DNA and RNA and establishing cell lines from target species will facilitate these initiatives. We provide a legal and methodological guide according to four standards of acquiring and storing tissue for the Genome 10K Project and similar initiatives as follows: four-star (banked tissue/cell cultures, RNA from multiple types of tissue for transcriptomes, and sufficient flash-frozen tissue for 1 mg of DNA, all from a single individual); three-star (RNA as above and frozen tissue for 1 mg of DNA); two-star (frozen tissue for at least 700 μg of DNA); and one-star (ethanol-preserved tissue for 700 μg of DNA or less of mixed quality). At a minimum, all tissues collected for the Genome 10K and other genomic projects should consider each species' natural history and follow institutional and legal requirements. Associated documentation should detail as much information as possible about provenance to ensure representative sampling and subsequent sequencing. Hopefully, the procedures outlined here will not only encourage success in the Genome 10K Project but also inspire the adaptation of standards by other genomic projects, including those involving other biota.
新测序技术在速度和效率上的最近提升促进了高通量测序、基因组组装和分析,推进了分析主要脊椎动物类群遗传序列的持续努力。从目标物种获取高质量 DNA 和 RNA 并建立细胞系的标准化程序将促进这些举措。我们根据获取和储存组织的四个标准,为 10K 基因组计划和类似的项目提供了法律和方法指南,如下所示:四星(保存的组织/细胞培养物、来自多种组织的转录组 RNA,以及来自单个个体的至少 1 毫克的 DNA 充足的速冻组织);三星(如上所述的 RNA 和至少 1 毫克的 DNA 冷冻组织);二星(至少 700μg 的 DNA 冷冻组织);一星(700μg 或更少混合质量的乙醇保存组织)。至少,为 10K 基因组和其他基因组项目收集的所有组织都应考虑每个物种的自然历史,并遵循机构和法律要求。相关文件应尽可能详细地说明来源信息,以确保代表性采样和后续测序。希望这里概述的程序不仅能鼓励 10K 基因组计划的成功,还能激励其他基因组项目,包括涉及其他生物群的项目,适应这些标准。