Yuan Yinan, SanMiguel Phillip J, Bennetzen Jeffrey L
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
Plant J. 2003 Apr;34(2):249-55. doi: 10.1046/j.1365-313x.2003.01716.x.
Higher eukaryotic genomes, including those from plants, contain large amounts of repetitive DNA that complicate genome analysis. We have developed a technique based on DNA renaturation which normalizes repetitive DNA, and thereby allows a more efficient outcome for full genome shotgun sequencing. The data indicate that sequencing the unrenatured outcome of a Cot experiment, otherwise known as High-Cot DNA, enriches genic sequences by more than fourfold in maize, from 5% for a random library to more than 20% for a High-Cot library. Using this approach, we predict that gene discovery would be greater than 95% and that the number of sequencing runs required to sequence the full gene space in maize would be at least fourfold lower than that required for full-genome shotgun sequencing.
包括植物基因组在内的高等真核生物基因组含有大量重复DNA,这使基因组分析变得复杂。我们开发了一种基于DNA复性的技术,该技术可使重复DNA标准化,从而使全基因组鸟枪法测序获得更高效的结果。数据表明,对Cot实验的未复性结果(即高Cot值DNA)进行测序,可使玉米中的基因序列富集四倍以上,从随机文库的5%增加到高Cot值文库的20%以上。使用这种方法,我们预测基因发现率将超过95%,并且对玉米全基因空间进行测序所需的测序运行次数将比全基因组鸟枪法测序所需的次数至少低四倍。