Plant Diversity ›› 2010, Vol. 32 ›› Issue (6): 535-538.DOI: 10.3724/SP.J.1143.2010.10069

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Effect of New Specificity and Yield Enhancer on Amplification of lea 3 Gene

 XU  Li-Xia, WU  Xiao-Lu, TANG  Xiao-Qian, YAN  Bo   

  1. Faculty of Landscape Architecture, Southwest Forestry University, Kunming 650224, China
  • Received:2010-04-01 Online:2010-12-25 Published:2010-07-02
  • Supported by:

    The Applied Foundation Research of Yunnan Province in China (Project number 2009ZC079M) and Yunnan Province and Minister Key Subject, University Key Lab and Share Platform

Abstract:

Amplification of a target DNA by the polymerase chain reaction (PCR) often requires experimental optimization factors especially when the target sequence contains high content of GC. Dimethyl sulfoxide, formamide, glycerol, NP40 and Tween20 are known to increase, the specificity and efficiency of PCR under certain conditions. In the amplification of lea3 gene in Gramineae, we found a new Extreme Thermostable SingleStranded DNA Binding Protein (ET SSB) and improved yield of PCR. Addition of 200 ng of ET SSB per 50 μl reation decreased the formation of nonspecific DNA fragments and increased the yield of specific PCR products.

Key words: High GC content, PCR, ET SSB

CLC Number: