Establishment and optimization of SRAP-PCR system in Canavalia ensiformis

Objective The purpose of this study is to establish the SRAP-PCR reaction system for Canavalia ensiformis.

Method An optimization experiment with single factor design was conducted, comprising five factors of Taq DNA polymerase, Mg²⁺, primer, dNTPs and DNA template, each with eight concentration levels, aiming to screen their suitable concentration range. After that, uniform design U₁₆(4⁵)and U₁₂(3⁵) were operated in order to improve the accuracy.

Result and conclusion The results showed that the optimum SRAP-PCR system was established, including Mg²⁺ 1.75 mmol/L, dNTPs 200 μmol/L, primers 0.36 μmol/L, Taq DNA polymerase 0.06 U/μL and DNA template 40 ng in the 25 μL reaction system. The reaction system is steady and dependable, which can be applied to the analysis of Canavalia ensiformis by SRAP.