热带作物学报2017，Vol.38Issue(7)：1311-1315,5.DOI:10.3969/j.issn.1000-2561.2017.07.020
海南黎药胆木PsbA-trnH-PCR体系的建立与优化
Establishment and Optimization of PsbA-trnH-PCR System for Li Nationality Medicine Nauclea officinalis Pierre ex Pitard of Hainan
摘要
Abstract
The study intended to establish a PsbA-trnH-PCR system and provide technical support to study the genetic diversity of Li Nationality medicine Nauclea officinalis Pierre ex Pitard in different geographical populations. High quality genomic DNA of N. officinalis Pierre ex Pitard was extracted by the plant genome kit method. The main factors in the process of PsbA-trnH-PCR were optimized by single factor and orthogonal design experiments, and PsbA-trnH-PCR products were sequenced. The results showed that the optimal PsbA-trnH-PCR reaction system (25μL) was Taq DNA polymerase 1.0 U, dNTPs 0.4 mmol/L, Mg2+ 0.75 mmol/L, primers 0.15μmol/L, template DNA 10 ng, 10×PCR Buffer (free Mg2+) 2.5μL. The genomic DNA of N. officinalis was amplified using the optimal PsbA-trnH-PCR system to obtain the amplification products, which were sequenced to obtain PsbA-trnH partial sequences. The stable PsbA-trnH-PCR system was established, and can be used in the identification and genetic diversity of for the study of N. officinalis.关键词
胆木/PsbA-trnH-PCR/优化系统/正交设计Key words
Nauclea officinalis Pierre ex Pitard/PsbA-trnH-PCR/optimization of system/orthogonal design分类
医药卫生引用本文复制引用
杨卫丽,高新征,邹园生,姚瑰玮,黄静,王勇..海南黎药胆木PsbA-trnH-PCR体系的建立与优化[J].热带作物学报,2017,38(7):1311-1315,5.基金项目
海南省自然科学基金(No. 20158282). （No. 20158282）
