WANG Xinmin1,ZHENG Renhua2,CHEN Jinhui1,ZHAO Yaqi1,XU Yang1,WANG Ying1,SHI Jisen1*
Author information+
1.Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education,Nanjing Forestry University, Nanjing 210037, China; 2. Fujian Academy of Forestry Sciences, Fuzhou 350012,China
In order to optimize SRAP-PCR reaction system for Chinese fir,the orthogonal design L16(45)was considered with different concentrations of Mg2+, dNTPs, primer, Taq DNA polymerase and DNA template in SRAP-PCR reaction system for Chinese fir. The results of PCR were evaluated by visual and variance analysis,then the concentrations of Mg2+,dNTPs,primer were rectified with single-factor design. Finally,the results showed that the best reaction system was the combination in 20 μL reaction solution,containing of Mg2+ 2.25 mmol/L,dNTPs 0.15 mmol/L,primer 0.4 μmol/L,Taq DNA polymerase1.5 μmol/min and DNA template 60 ng,2 μL10×PCR Buffer and ddH2O to 20 μL. The first optimal annealing temperature was 35 ℃,and the second step was 53 ℃. The optimized reaction system of Chinese fir described above can get clear and stable bands.
WANG Xinmin,ZHENG Renhua,CHEN Jinhui,ZHAO Yaqi,XU Yang,WANG Ying,SHI Jisen.
Optimization of Chinese fir SRAP-PCR system[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2014, 38(01): 15-20 https://doi.org/10.3969/j.issn.1000-2006.2014.01.003
中图分类号：
S722
Q81
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