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《Journal of Shanghai Jiaotong University (Agricultural Science)》 2002-01
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DNA extraction from single seeds and optimizing RAPD procedure in rice

PENG Suo tang,YAN Qi chuan,WANG Xue de,ZHANG Chun rong (Agricultural and Biotechnological Institute, Zhejiang University, Hangzhou 310029,China )  
By using single dry seeds of rice as experimenting materials, DNA extraction and optimum conditions of RAPD procedure were studied . The results showed that the effective amplification products could be obtained with DNA templates extracted from dry seeds by CTAB buffer. The optimum reaction system and conditions are: 100~400 μg/L random primers, 2~4 mmol/L Mg 2+ , 100~400 μmol/L dNTPs, 1 U Taq polymerase. Forty PCR cycles were performed, with 30s of denaturation at 94℃,30s of annealing at 38℃ and 1 min of polymerization at 72℃. To save time and money, predenaturation could be left out and the volume of the reaction may decrease to 12.5 μl.
【Key Words】: rice seed CTAB RAPD
【Fund】: 浙江省科技厅国际合作项目“中波种子质量改进与现代种子检测技术”资助 (1999-4 13 )
【CateGory Index】: S511.035.1
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