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《Journal of Beijing Forestry University》 2006-03
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Establishment and optimization of an ISSR reaction system for Populus tomentosa Carr.

FENG Xia-lian~1;HE Cheng-zhong~1;ZHANG Zhi-yi~1;AN Xin-min~1;YANG Kai~2;ZHANG You-hui~3.1 Key Laboratory for Genetic and Breeding in Forest Trees and Ornamental Plants, Ministry of Education,Beijing Forestry University,100083,P.R.China;2 Department of Biotechnology,Beijing Agricultural College,102206, P.R.China;3 Guanxian County Tree Nursery,Shandong Province,252528,P.R.China.  
In order to use ISSR markers to study genetic variation,assistant breeding,cultivar identification and phylogenesis of Populus tomentosa,the annealing temperature,concentration of primer,dNTP,Mg~(2+) and Taq DNA polymerase dosage on ISSR-PCR amplications were tested to determine their optimal levels and establish the following optimal reaction system for ISSR analysis in P.tomentosa.The results showed that the optimal conditions for ISSR-PCR of P.tomentosa were as follows: PCR reaction volume of 20 μL,1×Taq buffer(10 mmol/L Tris-HCl,50 mmol/L KCl,0.1% Triton X-100,pH 9.0),1.0 U Taq DNA Polymerase,0.2 μmol/L primer,0.2 mmol/L dNTP,1.5 mmol/L MgCl-2 and 10 ng template DNA.The optimized annealing temperature was 61℃ for primer(GTG)-6.
【Fund】: 国家自然科学基金项目(30571516)
【CateGory Index】: S792.117
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