中国水稻科学

中国水稻科学

• 实验技术 • 上一篇    下一篇

菌落PCR产物直接测序方法的建立及在水稻基因测序中的应用

毛伟华   

  1. 浙江大学 分析测试中心， 浙江 杭州 310029
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-09-10 发布日期:2005-09-10

Establishment of Direct Sequencing Method with Colony PCR Products and Its Application in Rice Gene Sequencing

MAO Wei-hua   

  • Received:1900-01-01 Revised:1900-01-01 Online:2005-09-10 Published:2005-09-10

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摘要： 建立了以菌落PCR 产物作为DNA 测序模板进行快速测序的技术方法。研究结果表明，采用菌落PCR技术，以载体插入位点两端序列互补的通用引物(如M13正反向引物)为引物，在控制菌落PCR反应条件的情况下,不仅可用于筛选和鉴定阳性克隆，而且菌落PCR 产物还可作为DNA 测序模板，结果准确可靠。与常规质粒测序方法比较，该法快速、简便，但对具有PolyT、PolyA过多的序列进行测序时，该法测序效率较低。

关键词: 菌落PCR, 质粒, 测序, 实验技术, 水稻

Abstract: A method for direct sequencing using colony PCR products was established.Under controlled conditions,colony PCR could be used to screen and identify positive clone,and its products could be used as sequencing templates directly.The sequencing result was reliable.Compared with conventional plasmid DNA sequencing method,colony PCR products sequencing method was faster and simpler.However,when the products were used to sequence the DNA with long PolyT and PolyA,the sequencing efficiency was low.

Key words: colony PCR, plasmid, sequencing, methodology, rice