Nature最新发表了一篇名为“The expanding scope of DNA sequencing”的一篇文章,讲述DNA测序技术的发展及其在各个领域中的应用在不断扩大。
短短的七年时间,下一代测序技术的成本降低,并且测序的速度也提高了4个数量级,常规的测序实验中都有数以百万计的测序结果。在研究中,测序不仅仅是用于组装基因组和调查人类疾病的遗传基础,同时也用于探索生物和细胞生物中的无数现象。在临床试验中,测序数据集中用于评估不同的研究背景,包括生殖医学,肿瘤和感染性疾病。新的测序应用的发展,一个反复出现的主题即根据实验标准进行的“重组”。但是,在下一代DNA测序的应用中仍然存在很多尚未可知的潜在影响。
最早的测序计划主要用于测定个人感兴趣的物种的基因组。随着新技术的出现,这些项目可以得到整个类群,甚至在这些类群家族中已经灭绝的物种的基因组。下一代测序技术为更精细的研究生物系统提供帮助。例如,我们能够探索单一物种中个别成员之间的遗传变异,和单个细胞之间的遗传与后天的差别。测序也提供了一个观察不同时期细胞的门径,包括所有显示出的现象。
文章更加形象的总结了各种不同的实验,这些实验最终都要进行测序,其中包括有8条实验线路,详细内容如下:
1、ChIP-Seq:Comparison, 'anatomic', cell culture, cross-linking, mechanical shearing, immunoprecipitation, DNA extraction, adaptor ligation, PCR and sequencing
2、TRAP:Comparison, genetic manipulation, 'anatomic', cross-linking, affinity purification, RNA extraction, poly(A) selection, reverse transcription, second-strand synthesis, adaptor ligation, PCR and sequencing
3、RNA-Seq:Comparison, 'anatomic', RNA extraction, poly(A) selection, chemical fragmentation, reverse transcription, second-strand synthesis, adaptor ligation, PCR and sequencing
4、Hi-C:Comparison, cell culture, cross-linking, proximity ligation, mechanical shearing, affinity purification, adaptor ligation, PCR and sequencing
5、Immuno-Seq:Perturbation, 'anatomic', DNA extraction, PCR and sequencing
6、Methyl-Seq:Perturbation, genetic manipulation, cell culture, DNA extraction, mechanical shearing, adaptor ligation, bisulfite conversion, PCR and sequencing
7、Synthetic saturation mutagenesis:Variation, genetic manipulation, barcoding, RNA extraction, reverse transcription, PCR and sequencing
8、Exome-Seq:Comparison, 'anatomic',Toal DNA extraction,Hybrid capture,PCR and sequencing
原文摘要:
The expanding scope of DNA sequencing
Jay Shendure, Erez Lieberman Aiden
In just seven years, next-generation technologies have reduced the cost and increased the speed of DNA sequencing by four orders of magnitude, and experiments requiring many millions of sequencing reads are now routine. In research, sequencing is being applied not only to assemble genomes and to investigate the genetic basis of human disease, but also to explore myriad phenomena in organismic and cellular biology. In the clinic, the utility of sequence data is being intensively evaluated in diverse contexts, including reproductive medicine, oncology and infectious disease. A recurrent theme in the development of new sequencing applications is the creative 'recombination' of existing experimental building blocks. However, there remain many potentially high-impact applications of next-generation DNA sequencing that are not yet fully realized.