松江鲈鱼ISSR-PCR反应体系的建立及优化研究(英文)

[Objective] The aim of this study is to establish and optimize the ISSR-PCR system in roughskin sculpin(Trachidermus fasciatus Heckel).[Method] By using the primer of ISSR-63,the concentrations of Taq DNA polymerase,Mg2+,dNTPs and primer were optimized by orthogonal design for establishing the suitable ISSR-PCR system in roughskin sculpin;moreover,the suitable anneal temperature was yielded from gradient PCR on temperature.[Result]The optimized ISSR-PCR system(20 μl reaction volume)in roughskin sculpin was proved to be:2.5 mmol/L Mg2+,250 μmol/L dNTPs,0.25 μmol/L primer,1 U Taq DNA polymerase,30 ng DNA template and 1×PCR buffer;and suitable anneal temperature was determined to be 50.8 ℃.The established system was further confirmed by using 24 wild roughskin sculpin samples.[Conclusion]The results lay a foundation for the analysis of genetic diversity and germplasm resources of roughskin sculpin.

Establishment and Optimization of ISSR-PCR System in Trachidermus fasciatus Heckel

[Objective] The aim of this study is to establish and optimize the ISSR-PCR system in roughskin sculpin (Trachidermus fasciatus Heckel). [Method] By using the primer of ISSR-63, the concentrations of Taq DNA polymerase, Mg2+ , dNTPs and primer were optimized by orthogonal design for establishing the suitable ISSR-PCR system in roughskin sculpin; moreover, the suitable anneal temperature was yielded from gradient PCR on temperature. [Result] The optimized ISSR-PCR system(20 μl reaction volume) in roughskin sculpin was proved to be: 2.5 mmol/L Mg2+, 250 μmol/L dNTPs, 0.25 μmoL/L primer, 1 U Taq DNA polymerase, 30 ng DNA template and 1×PCR buffer; and suitable anneal temperature was determined to be 50.8 ℃. The established system was further confirmed by using 24 wild roughskin sculpin samples. [Conclusion] The results lay a foundation for the analysis of genetic diversity and germplasm resources of roughskin sculpin.