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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (31): 93-98.

• 林学 园艺 园林 • 上一篇    下一篇

绣球属植物SRAP-PCR反应体系优化及引物筛选

陈海霞 彭尽晖   

  • 收稿日期:2011-05-30 修回日期:2011-07-25 出版日期:2011-12-05 发布日期:2011-12-05

Optimization of SRAP-PCR System and Primers Screening in Hydrangea

  • Received:2011-05-30 Revised:2011-07-25 Online:2011-12-05 Published:2011-12-05

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摘要：

为了确定绣球属植物SRAP-PCR最适宜的反应体系，以19种绣球属植物为材料，利用单因素分析法对影响SRAP-PCR反应体系的5个因素（DNA模板量，Mg2+浓度，dNTPs浓度，Taq聚合酶量和引物浓度）在11个水平上进行优化试验。结果表明，最佳的25 μL反应体系为：10×PCR Buffer 2.5 μL，DNA模板量30 ng，Mg2+浓度1.6 mmol/L、dNTPs浓度0.6 mmol/L，Taq聚合酶量3.5 U，引物浓度为0.2 μmol/L。单因素分析法获得的最佳反应体系适合绣球属植物SRAP的遗传多样性研究。

关键词: 保水剂、绿豆、生理性状、产量, 保水剂、绿豆、生理性状、产量

Abstract:

In order to make clear the optimizing SRAP-PCR reaction system of Hydrangea, 19 Hydrangea plants were used as material, the single factor experiment was designed in 11 levels of 5 factors (concentration of primer, Mg2+, DNA template and dNTPs, and Taq DNA polymerase contents) respectively. The results showed that the reaction system obtained as followed: 2.5 μL 10×PCR Buffer, 30 ng DNA, 1.6 mmol/L MgCl2, 0.6 mmol/L dNTPs, 3.5 U Taq DNA polymerase, and in a total volume of 25 μL. This optimizing reaction system is suit for the SRAP genetic variation research of Hydrangea.