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小麦不同部位DNA提取的比较研究

         

摘要

[目的] 探讨小麦不同取材部位对DNA提取质量和产率的影响.[方法] 分别以小麦的种子、根尖和幼嫩叶片为材料,采用改良CTAB法分别提取小麦3个部位的基因组DNA,用0.8%的琼脂糖凝胶电泳检测DNA质量,用紫外分光光度计分别测定3个部位的DNA在A260和A280下的吸光值,根据A260/A280的值检测DNA的浓度和纯度,并以3个部位提取的DNA为模版进行RAPD扩增.[结果] 采用小麦叶片提取DNA的产率最高,种子次之,根尖最低.3个部位提取的DNA纯度均相近, 提取叶片获得的DNA浓度最大,为 756 μg/ml;其次是种子,为233 μg/ml;根尖获得的DNA浓度最低,为90 μg/ml.小麦不同部位提取的DNA模板对RAPD扩增没有明显影响,DNA浓度都能达到扩增的要求.[结论] 小麦3个部位提取的DNA质量均较好,可以进行后续的酶切和PCR扩增等实验.%[Objective] The aim was to discuss the effect of different sampling positions of wheat on the DNA quality and yield. [Method] With seeds, root tips and young leaves of wheat as tested materials, the genome DNA from 3 parts of wheat were extracted with modified CTAB method and examined with 0.8% agarose gel electrophoresis. The absorbance value of DNA from 3 parts was measured by ultraviolet spectrophotometer at A260 and A280 and the ratio of A260/A280 was used to determine the DNA purity and the mass concn. DNA isolated from 3 parts of wheat was used as the template for RAPD amplification. [Result] The yield of DNA extracted from wheat leaves was highest, that from the seeds was second, and that from the root tips was lowest. The DNA purity extracted from 3 parts of wheat was similar. The DNA concn. isolated from wheat leaves was the largest, being 756 μg/ml, followed by that from seeds, being 233 μg/ml that from the root was the lowest, being 90 μg/ml. The DNA template extracted from different parts of wheat had no significant effect on RAPD amplification and DNA concn. could achieve the amplification requirements. [Conclusion] The DNA qualities extracted from 3 parts of wheat were good and could be used in subsequent experiments such as digestion and PCR amplification.

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