Optimization and Verification of SRAP-PCR System for Culinary Rhubarb

doi:10.7668/hbnxb.2014.04.017

Abstract

Abstract: The purpose of this study was to determine the best system of SRAP reaction and provide a foundation for further studying the genetic diversity of culinary rhubarb.Using the genome DNA as template,and combining the single-factor experiment with the orthogonal design,five factors including the concentrations of template DNA,Mg²⁺,dNTPs, Taq DNA polymerase and primers in SRAP-PCR reaction were optimized,and then the optimal SRAP-PCR reaction system for culinary rhubarb was established.The results indicated that the order of each factor affecting the result of PCR was:primer>Mg²⁺>dNTPs> Taq DNA polymerase dosage>DNA.The optimal SRAP-PCR reaction for culinary rhubarb in a 25 μL reaction system was 10×PCR Buffer 2.50 μL,template DNA 30 ng,Mg²⁺ 2.50 mmol/L,dNTPs 0.25 mmol/L, Taq DNA polymerase dosage 0.04 U/μL and each primer 0.5 μmol/L.The optimized system was verified by the genomic DNA samples from eight cultivars of culinary rhubarb,and the clear bands and abundant polymorphism were obtained.It was concluded that the SRAP-PCR reaction system was stable and reliable,and was suitable to analyze the genetic diversity of culinary rhubarb.

Key words: Culinary rhubarb, SRAP-PCR, Optimization

CLC Number:

S644.9

GUO Xiao-fei, JIANG Li-na, CAI Zu-guo, REN Wen-juan, ZHAO Yi-peng. Optimization and Verification of SRAP-PCR System for Culinary Rhubarb[J]. journal1, 2014, 29(4): 105-110. doi: 10.7668/hbnxb.2014.04.017.

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http://www.hbnxb.net/EN/Y2014/V29/I4/105

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