福建林学院学报Issue(2)：165-170,6.
凹叶厚朴 TRAP-PCR 体系建立
Establishment of TRAP-PCR system for Magnolia officinalis subsp.biloba
摘要
Abstract
Uniform design and orthogonal design methods were applied in this paper to optimize 5 key factors ( template DNA, Mg2+, dNTPs, primers and Taq polymerase) that could influence the effect of TRAP-PCR.The TRAP-PCR system was set up in Magnolia officinalis subsp.biloba.The optimal system is in 25 μL reaction volume containing about 40 ng template DNA , 1.5 mmol· L-1 of Mg2+, 0.28 mmol· L-1 of dNTPs, 0.3 μmol· L-1 of fixed primer and arbitrary primer , and 0.75 U Taq DNA polymerase.PCR program:predenaturing 5 min at 94 ℃; the first five cycles run at 94 ℃, 1 min, 40 ℃, 1 min, and 72 ℃, 1 min, for denaturing , annealing and extension , respectively; then the annealing temperature is raised to 50 ℃ for another 38 cycles;run 10 min at 72 ℃for extension .Using the optimization system for amplification of 16 different provenances of M.officinalis subsp.biloba, results showed that the system had good effect and could be used in the correlational studies of M.officinalis subsp.biloba.关键词
凹叶厚朴/TRAP/均匀试验/正交试验/优化Key words
Magnolia officinalis susbsp.biloba/TRAP/uniform design/orthogonal design/optimization分类
农业科技引用本文复制引用
翁剑,李单琦,李书平,荣俊冬,周少卿,郑郁善..凹叶厚朴 TRAP-PCR 体系建立[J].福建林学院学报,2014,(2):165-170,6.基金项目
福建省科技重点基金项目（2009N003）；国家“十二五”科技支撑项目（2011BA101B06）。 （）
