枣属ITS及psbA-trnH序列PCR反应体系的优化与验证

Optimization and Verification of PCR Reaction System for ITS and psbA-trnH Sequences Amplification of Ziziphus Mill.

为了建立和优化枣属植物核rDNA的ITS及cpDNA的psbA-trnH序列PCR反应体系,采用正交试验和单因子试验方法,对影响PCR扩增体系中的DNA模板、退火温度2个重要因子进行了比较分析。结果表明:枣属50滋L反应体系中,DNA模板量为50~200ng时,不同的DNA模板量对PCR反应体系影响不大;退火温度对PCR反应影响较为明显,最佳退火温度为52℃,扩增时psbA-trnH片段要求的退火温度较ITS序列更为严格。

To establish and optimize the PCR reaction system for ITS and psbA-trnH sequences of Ziziphus Mill.,using orthogonal test and single factor test,the template DNA and annealing temperature were compared. The results showed that 50-200 ng DNA in 50℃L PCR reaction system had little influence on PCR amplification,while the annealing temperature had obvious effect,and the optimum annealing temperature was 52 益. The annealing temperature for amplification of psbA-trnH sequence should be controlled more strictly than that of ITS sequence.