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利用CTAB法、SDS法、Trizol法,从绿豆叶片中提取了总RNA;利用改进的GT法从绿豆叶、茎和子叶中提取了总RNA,提取结果通过凝胶电泳和紫外吸光光度值检测。结果显示:用改良的GT法提取的总RNA,凝胶电泳图上28S条带是18S条带亮度的两倍以上;而用其它3种方法提取的总RNA电泳条带不如GT法提取的总RNA清晰。用GT法从绿豆叶片中提取的总RNA的A260/A280值为1.94,可以用于RT-PCR、DDRT-PCR和Northern杂交等实验。通过RT-PCR,得到绿豆钙调蛋白基因约450bp,从而进一步检验了提取的总RNA的质量。改良的GT法可以认为是一种简便、快速的提取绿豆RNA的方法。
Total RNA was extracted from mung bean leaves by CTAB method, SDS method and Trizol method. Total RNA was extracted from mung bean leaves, stems and cotyledons using improved GT method. The results were analyzed by gel electrophoresis and UV spectrophotometry. The results showed that the total RNA extracted by the modified GT method showed that the 28S band on the gel electrophoresis pattern was more than twice the brightness of the 18S band. However, the total RNA bands extracted by the other three methods were not as good as those obtained by the GT method Clear RNA. The A260 / A280 value of total RNA extracted from mung bean leaves by GT method was 1.94 and could be used for experiments such as RT-PCR, DDRT-PCR and Northern hybridization. By RT-PCR, mung bean calmodulin gene was obtained about 450bp, so as to further test the quality of extracted total RNA. The modified GT method can be considered as a simple and rapid method for extracting mung bean RNA.