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| The effects of several amplification conditions on RAPD amplification for Alsophila spinulosa |
| LI Xue-yan1, WANG Ting2, SU Ying-juan2 |
| 1. Department of Biotechnology of Zhongshan College, University of Electronic Science and Technology of China, Zhongshan 528402; 2. School of Life Sciences, Zhongshan University, Guangzhou 510275, China |
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Abstract To get the optial amplification conditions for Alsophila spinulosa,the effects of the purified methods of DNA templates and the concentrations of DNA templates,Taq DNA polymerase,magesium,primer and dNTP are tested.The results suggest:the purified templates by glass milk or proteinase K are brighter,clearer and more repeatable than the unpurified templates or precipitated templates by ethanol;the concentration of DNA templates,Taq DNA polymerase,magesium,primer and dNTP will influence the productions of RAPD.The optimal amplification conditions for Alsphila spinulosa is acquired through the experiments:the concentration of DNA templates is 70ng,the concentration of magnesium is 2.5 mmol·L-1,dNTP is 0.2 mmol·L-1,primer is 3×104 mL.The optical amplification program is also obtained:94·200s,1 cycle;94℃ 60s,36℃ 60s;72℃ 120s,40 cycle;72℃ 600s,1 cycle;stored in 4℃.
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Received: 25 April 2003
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