基于四段基因序列的野牡丹科植物PCR反应条件优化

Optimization of PCR Reaction Conditions of Melastomataceae Based on Four Gene Region

为了建立植物分子系统学研究中常用的核基因的ITS,CHS和叶绿体基因的psbA-trnH,rpl16序列在野牡丹科植物中的PCR扩增反应的最佳反应体系,寻找最佳反应条件。采用改良的试剂盒法提取了15种野牡丹科植物的基因组DNA,进行了引物筛选,退火温度优化,添加牛血清蛋白(BSA)和二甲基亚砜(DMSO)的优化,反应体系正交设计一系列优化。并采用优化后的反应条件,对这4个基因片段在15种野牡丹科植物中进行PCR扩增验证。结果表明:优化后的反应条件在野牡丹科植物中具有较高的稳定性和良好的通用性。说明优化后的反应条件可以进行后续分子系统学研究。

In order to establish the suitable PCR amplification system and find the best conditions of nr-ITS,nr- CHS,cp-psbA-trnH and cp-rpl16, which are frequently used in Plant Molecular Phylogeny research, after the genomic DNA of 15 species in Melastomataceae was extracted by modified Kit method,a series of optimization, such as primer screening, annealing temperature optimization, add BSA and DMSO, and reaction-system orthog- onal design were made. 15 species of Melastomataceae were used to test the reaction conditions after optimization. The results indicated that the optimized reaction conditions were suitable and stable for the PCR amplification of Melastomataceae. It showed that the reaction conditions after optimization can provides reference for subsequent molecular phylogeny research.