期刊论文

中文

西北农业学报

1004-1389

2008

17

6

148-152,183

10.3969/j.issn.1004-1389.2008.06.032

本校为第一机构

利用L45正交设计对影响南瓜SRAP反应体系的MgCl2浓度、dNTPs浓度、Taq酶含量、引物浓度及模板DNA浓度等5个因素进行了筛选，快速得到稳定性和重复性好的南瓜SRAP扩增体系。对复性温度、PCR循环次数等影响南瓜SRAP扩增结果的重要因素进行了优化。最终优化的南瓜SRAP反应体系为25μL反应液中：10×buffer2.5μL，0.2 mmol/L dNTPs，引物0.2μmol/L，1.5 mmol/LMgCl2，DNA模板6 ng/μL，Taq酶1.5 U。本研究最终确立的PCR反应程序为：94℃预变性5 min，94℃变性40 s，35℃退火40 s，72℃延伸90 s，进行5个循环，94℃变性40 s，50℃退火40 s，72℃延伸90 s，进行32个循环，最后72℃延伸5 min。在此条...

The optimum reaction system of SRAP in squash was studied in order to ensure stability and product bility of SRAP. After testing some important influencing factors of SRAP in squash such as MgCl2 ,dNTPs, Taq DNA polymerase, primer, template DNA, the temperature for annealing and the number of cycles,the results showed that in 25 uL SRAP reaction system , the optimum concentration were 1.5 mmol/L MgCl2, 0. 2 mmol/L dNTPs, Taq DNA polymerase 1. 5 U, primer 0.2 ng/uL, template DNA 6 ng/uL. The PCR amplification procedure used in this study was as ...