相關(guān)專題

相關(guān)專題：測序

Nature發(fā)表了一篇名為“The expanding scope of DNA sequencing”的一篇文章，講述DNA測序技術(shù)的發(fā)展及其在各個領(lǐng)域中的應(yīng)用在不斷擴大。

短短的七年時間，下一代測序技術(shù)的成本降低，并且測序的速度也提高了4個數(shù)量級，常規(guī)的測序?qū)嶒炛卸加袛?shù)以百萬計的測序結(jié)果。在研究中，測序不僅僅是用于組裝基因組和調(diào)查人類疾病的遺傳基礎(chǔ)，同時也用于探索生物和細胞生物中的無數(shù)現(xiàn)象。在臨床試驗中，測序數(shù)據(jù)集中用于評估不同的研究背景，包括生殖醫(yī)學(xué)，腫瘤和感染性疾病。新的測序應(yīng)用的發(fā)展，一個反復(fù)出現(xiàn)的主題即根據(jù)實驗標準進行的“重組”。但是，在下一代DNA測序的應(yīng)用中仍然存在很多尚未可知的潛在影響。

zui早的測序計劃主要用于測定個人感興趣的物種的基因組。隨著新技術(shù)的出現(xiàn)，這些項目可以得到整個類群，甚至在這些類群家族中已經(jīng)滅絕的物種的基因組。下一代測序技術(shù)為更精細的研究生物系統(tǒng)提供幫助。例如，我們能夠探索單一物種中個別成員之間的遺傳變異，和單個細胞之間的遺傳與后天的差別。測序也提供了一個觀察不同時期細胞的門徑，包括所有顯示出的現(xiàn)象。

文章更加形象的總結(jié)了各種不同的實驗，這些實驗zui終都要進行測序，其中包括有8條實驗線路，詳細內(nèi)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.