青檀SRAP-PCR体系优化设计方案

以青檀(Pteroceltis tatarinowii Maxim.)叶片为材料,采用正交设计和均匀设计两种方法对SRAP-PCR反应体系进行优化,并对这两种设计方案优化出的最佳反应体系进行比较,结果表明:两种设计均可用于青檀SRAP-PCR体系的优化,但与正交设计相比,均匀设计在多因素多水平条件下,得到的条带更清晰、稳定性更好。通过实验比较筛选出的青檀SRAP-PCR最佳反应体系为:2.5μL 10×PCR buffer,20 ng模板DNA,Mg2+2.5 mmol/L,dNTP150μmol/L,引物0.2μmol/L,Taq DNA聚合酶1.0 U,总体积25μL。

Comparison between orthogonal design and uniform design to optimize the reaction system for SRAP of Pteroceltis tatarinowii Maxim.

Orthogonal design and uniform design were used to obtain＇optimum scheme on reaction system for sequence-related amplified polymorphism （SRAP） of Pteroceltis tatarinowii Maxim. , and the optinmm reaction systems of the two designs were compared. The resuits showed that both orthogonal design and uniform design could be used for sequence-related amplified polymorphism of Pteroceltis tatarinowii. But compared with the orthogonal design, uniform design could quickly obtain the optimum reaction system for sequencerelated amplified polymorphism of Pteroceltis tatarinowii with much clearer bands and better stability. The optimal 25 txL reaction system of SRAP for Pteroceltis tatarinowii included 2.5 μL 10× PCR buffer, 20 ng template DNA, Mg^2＋2.5retool/L, dNTP 150μmol/L, primer 0.2μmol/L, Taq DNA polymerase 1. 0U.