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《Chinese Bulletin of Entomology》 2007-04
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Extraction of genomic DNA and optimization of RAPD system for Gomphocerus sibiricus

GE Feng-Wei1,JI Rong1,ZENG Xian-Chun1,WANG Ting1,WANG Xiao-Ping2(1.College of Life and Chemisty,Xinjiang Normal University,Urumqi 830054,China;2.Institute of Agricultural Professional Technology,Changji 831100,China)  
By using an improved SDS method,we obtained high qualified genomic DNA of Gomphocerus sibiricus(L.).The effects of several parameters(contents of dNTP,Mg2+,Taq DNA polymerase and template DNA) on amplification were examined.The optimal reaction system of RAPD of G.sibiricus was determined as followed:0.24 mmol/L dNTP,2.0 mmol/L Mg2+,1.0 U Taq DNA polymerase,45 ng template DNA and 30 ng primer in total 25 μL reaction volume.Modified thermal profile consisted of an initial denaturation step at 94℃ for 1 min and 45 s,followed by 45 cycles of 94℃ for 30 s,35℃ 1 min and 30 s and 72℃ for 2 min,and a final exposure to 72℃ for 10 min.
【Fund】: 国家自然科学基金项目(30460028);; 新疆维吾尔自治区高校科研资助项目(XJEDU2005I23 XJEDU2004S20)
【CateGory Index】: Q966
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