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Establishment of TRAP-PCR system for Magnolia officinalis subsp.biloba
Author(s): WENG Jian, LI Dan-qi, LI Shu-ping, RONG Jun-dong, ZHOU Shao-qing, ZHENG Yu-shan
Pages: 165-
170
Year: 2014
Issue:
2
Journal: Journal of Fujian College of Forestry
Keyword: Magnolia officinalis susbsp.biloba; uniform design; orthogonal design; optimization;
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.
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