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《Journal of Fujian Forestry Science and Technology》 2006-01
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Optimization of the ISSR-PCR reaction system of Quercus glandulifera var.brevipetiolata

DING Li-ya~(1,2),JIN Zen-xin~2,LI Jun-min~2(1.Beijing Forestry University College of Natural Resources and Enviroment,Beijing 100083,China;2.Ecology Institute of Taizhou University,Zhejiang Linhai 317000,China)  
Factors which affect the ISSR-PCR amplification such as template DNA usage,Taq DNA polymerase,Mg~(2+) concentration,dNTP concentration,primer usage and bovine serum albumin concentration,were optimized and selected by using the materials of Quercus glandulifera var.brevipetiolata DNA.The results showed that the conditions suitable for ISSR-PCR of Quercus glandulifera var.brevipetiolata were as follows:1×Taq buffer(10 mmol·L~(-1) Tris-HCl,pH9.0,50 mmol·L~(-1) KCl,0.1% Triton X-100),0.75U Taq DNA polymerase,2.0 mmol·L~(-1) MgCl_2,0.2 mol·L~(-1) 4×dNTP,12 pmol primers,10ng template DNA、2 mg·mL~(-1) bovine serum albumin. The suitable annealing temperature in the ISSR-PCR reaction system was 54.4℃.
【Fund】: 浙江省自然科学基金资助项目(Y504220);; 台州市科技局资助项目(044205)
【CateGory Index】: Q943.2
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