摘要
本文用水稻单粒干种子作材料 ,进行了 DNA提取方法和 RAPD程序优化研究。结果表明 :CTAB法微量提取水稻干种子 DNA可获得理想的扩增结果 ,最佳的反应体系和条件是 :随机引物浓度 1 0 0～ 40 0 μg/L、Mg2 + 浓度 2～ 4mmol/L、d NTPs浓度 1 0 0～ 40 0 μmol/L、Taq酶 1 U。变性 94℃ 3 0 s,退火 3 8℃ 3 0 s,延伸 72℃ 1 min,40个循环。为了节约时间和成本 ,可不进行予变性处理 ,反应体积可降至 1 2 .5μl,扩增效果一样。
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.
出处
《上海交通大学学报（农业科学版）》
2002年第1期34-37,41,共5页
Journal of Shanghai Jiaotong University(Agricultural Science)
基金
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 )
