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Journal of Tea Science ›› 2004, Vol. 24 ›› Issue (3): 172-176.doi: 10.13305/j.cnki.jts.2004.03.005

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Establishment of ISSR-PCR Reaction Conditions in Tea Plant

YAO Ming-zhe, WANG Xin-chao, CHEN Liang, YANG Ya-jun   

  1. Tea Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Tea Chemical Engineering, Ministry of Agriculture, Hangzhou 310008,China
  • Online:2004-09-25 Published:2019-09-16

Abstract: The optimal ISSR-PCR reaction conditions in tea was established by studying the main parameters. Results showed that the optimum concentration of five important components i.e. template DNA, primer, Mg++, dNTP, Taq DNA polymerase in 20βµl reaction mixture was 10ng, 150βnmol/L, 1.5βmmol/L, 150βµmol/L, 0.5βU, respectively. The appropriate annealing temperature was average 4.5℃ higher than Tm of corresponding primer, and at least 30 PCR cycles should be carried out to ensure sufficient PCR products. ISSR polymorphism between thirteen tea germplasm was 77.6%, and 12 out of 13 tea germplasm could be identified by ISSR fingerprinter established with primer TRI18.

Key words: Tea plant, ISSR, PCR reaction conditions

CLC Number: