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《Journal of Fujian Forestry Science and Technology》 2008-01
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The optimization of ISSR-PCR conditions of Liquidambar formosana

BI Quan-xin1,JIN Ze-xin1,ZENG Zhi-cheng1,YE Wen-guo2,LI Jian-hui1 (1.Ecological Research Institute of Taizhou College,Linhai Zhejiang 317000,China;2.Tiantai County Forestry Bureau,Tiantai,Zhajiang 317200,China)  
The optimal conditions of ISSR amplification of Liquidambar formosana from Tiantai Mountain in Zhejiang Province was screened.After the analysis of the concentration Mg2+,4×dNTP,BSA,template DNA dosage,primer dosage,and unit of Taq DNA polymerase,the optimal conditions of ISSR-PCR was determined as follows: 1×Taq buffer(10 mmol·L-1 Tris-HCl,pH9.0,50 mmol·L-1 KCl and 0.1% Triton X-100),2.0 mmol·L-1 Mg2+,0.15 mol·L-1 4×dNTP,2.0 mg·mL-1 BSA,10 ng template DNA,6 pmol primers and 0.9 U Taq DNA polymerase;the suitable annealing temperature of UBC808 primer in the ISSR-PCR reaction system was 52.4℃.Using the optimal amplification,91 loci were produced in L.formosana population of Tiantai Mountain.The polymorphic loci percentage was 84.26%,Nei's gene diversity was 0.2665,and Shannon's information index was 0.4044.The genetic diversity of L.formosana was in a high level.
【Fund】: 浙江省自然科学基金资助项目(Y504220)
【CateGory Index】: S792.99
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