欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (19): 94-99.

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

茶树cDNA-AFLP银染技术体系的建立

吴扬 邓婷婷 李娟 欧秋良 黄建安   

  • 收稿日期:2011-05-16 修回日期:2011-06-12 出版日期:2011-08-05 发布日期:2011-08-05
  • 基金资助:

    茶氨酸生物合成的基因转录调控研究

Establishment of cDNA-AFLP Sliver Staining Technical System in Tea Plant (Camellia sinensis)

  • Received:2011-05-16 Revised:2011-06-12 Online:2011-08-05 Published:2011-08-05

PDF (PC)

1157

摘要：

为了挖掘安白茶白化期相关的差异基因，建立并优化了茶树叶片cDNA-AFLP扩增反应体系，进行cDNA-AFLP分析。在安吉白茶叶片cDNA-AFLP的试验中，分别对20 μL预扩增和选择性扩增反应体系中的4种反应条件影响因子进行不同梯度优化的研究，包括引物、Mg2+、dNTP及Taq DNA聚合酶的用量。PCR预扩增20 μL反应体系中，引物75 ng/20 μL，Mg2+ 2.0 mM，dNTP 0.2 mM，Taq DNA聚合酶1 U时预扩增效果较好；PCR选择性扩增20 μL反应体系中，引物75 ng/20 μL，Mg2+ 2.5 mM，dNTP 0.2 mM，Taq DNA聚合酶1.5 U时选择性扩增效果较好。通过试验，获得较为理想的预扩增和选择性扩增体系。

关键词: 安全性, 安全性

Abstract:

In order to obtain white period leaves genes in Anji Bacha, cDNA-AFLP amplification reaction system was established and optimized, and cDNA-AFLP was analyzed. In the experiment of leaf in Anji Bacha, the optimization of cDNA-AFLP reaction system was studied by employing white and green period leaves as samples. Different levels of concentration of primer, dNTP mixture, Mg2+ and Taq DNA polymerase were studied in 20 μL amplification system. The results showed that amplification effects are good when the volume of primer concentration was 75 ng/20 μL, Mg2+ concentration was 2.0 mM, dNTP concentration was 0.2 mM, Taq DNA polymerase was 1 U in 20 μL pre-amplification system and the volume of primer concentration was 75 ng/20 μL, Mg2+ concentration was 2.5 mM, dNTP concentration was 0.2 mM, Taq DNA polymerase was 1.5 U in 20 μL selective amplification system. This test obtained better pre-amplification and selective amplification system.