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Molecular Biology

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Studying DNA and RNA

DNA Sequencing Methods and Uses

Description

DNA sequencing methods can read millions of DNA fragments at the same time. Next-generation sequencing made this process much faster and much less expensive than older methods. The first human genome project cost $2.7 billion and took 15 years. It was completed in 2003 after major international cooperation among research tea...

Transcript

下一代测序是一组技术的统称,可相对快速地从许多不同物种或个体群体中生成序列数据。

这些数据可用于精细尺度的群体遗传学或临床研究,且这些技术比传统测序方法更快、更具成本效益。

目前有许多下一代测序方法,但其中最流行的是可逆终止子测序法。其他方法包括焦磷酸测序法和连接测序法。

在可逆终止子测序中,首先将纯化的基因组DNA切割成100-1000个碱基对的小片段。然后通过连接将带有引物结合位点的特殊接头——即寡核苷酸序列——连接到这些DNA片段的两端。

随后将单链DNA片段加载到一种称为流动池的特殊芯片上,该芯片预先包被有与接头序列互补的短寡核苷酸序列。DNA片段通过其接头与芯片结合。

在扩增阶段,使用DNA聚合酶合成互补的...

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