Optimization of RFLP-PCR Reaction System for cpDNA in Tea Plants

doi:10.7506/spkx1002-6630-201306016

Abstract

Abstract: Optimal reaction system of PCR-restriction fragment length polymorphism (RFLP) for cpDNA in tea plants was investigated by orthogonal array design. Results showed that the optimal amplification system was 100 ng DNA template, 200 μmol/L dNTPs, 1.5 mmol/L MgCl2, 50 ng primer, 3U Taq DNA polymerase, and addition of ddH2O up to total volume of 25 μL. The optimal digestion system included 6 μL amplified product, 2 U endonuclease, 1 × endonuclease buffer in digestion solution, digestion time of 6 h and digestion temperature of 37 ℃, and addition of ddH2O up to total volume of 15 μL. Under the optimal reaction system, 30 camellia cultivars were analyzed and polymorphic digestion maps were obtained.

Key words: tea plant, chloroplast DNA, PCR-RFLP (restriction fragment length polymorphism), optimization

CLC Number:

S571.1

CHEN Sheng-xiang，QI Gui-nian*，LI Huan. Optimization of RFLP-PCR Reaction System for cpDNA in Tea Plants[J]. FOOD SCIENCE, 2013, 34(6): 73-76.

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