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Bulletin of Botanical Research ›› 2006, Vol. 26 ›› Issue (3): 318-322.doi: 10.7525/j.issn.1673-5102.2006.03.014

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Optimization of experiment conditions and primer screening with ISSR markers for Amentotaxus argotaenia

ZHU Bai-Fang;ZHU Du*;DENG Rong-Gen;LI Si-Guang   

  1. 1.College of life science, Jiangxi normal University/Jiangxi Provincial key lab of protection and utilization of subtopic plant resources, Nanchang 330027 2.College of life science, Nanchang University, Nanchang 330047
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-05-20 Published:2006-05-20
  • Contact: ZHU Du
  • Supported by:

Abstract: The factors which affect the ISSR analysis in the study of the genetic deversity of Amentotaxus argotaenia, such as concentrations of template DNA, DNA Taq polymerase,Mg2+ and dNTPsn et al., were examined. The optimal ISSR-PCR conditions in the experiments were as the following: in 20 μL PCR reaction system,2 μL 10×DNA Taq polymerase buffer, 1.8 U Taq DNA polymerase, 0.2 μmol·L-1 primer, 0.18 mmol·L-1 dNTP, 1.5~2.5 mmol·L-1 MgCl2, 10 ng·μL-1 temper DNA. One hundred ISSR primers were used to screen the suitable primers with 7 samples from different populations for assessing the genetic diversity of Amentotaxus argotaenia, of which 10 ISSR primers with high resolution and multiple polymorphic bands were screened. The total 92 ISSR bands were amplified with 10 primers, and produced 45 polymorphic bands. The percentage of polymorphic bands is 49%.

Key words: ISSR, primer screening, Amentotaxus argotaenia

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