不同方法提取绿豆总RNA的比较和改进

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。

Comparison and Improvement of Different Methods of RNA Isolation from Phaseolus radiatus L.

Basing on the methods of CTAB, SDS, Trizol, and improved GT, total RNA was isolated from leaves ofPhaseolus radiatus L.. The total RNA were compared by gel electrophoresis and the ratio of A260/A280. The result indicated that the quality of RNA was really high, and the sharpness of 28S bands was twice than 18S of bands by using GT method. While the qualities of RNA isolated by other methods were less than that GT method. A260/A280 of RNA isolated by GT was 1.94, the ratio is good enough for RT-PCR, DDRT-PCR and Northern blotting. After RT-PCR, the 450bp sequence ofcalmodulin gene is obtained to identify the quality of total RNA from P. radiatus L.. In this experiment, improved GT method can be regarded as a simple, rapid and convenient method for the RNA isolated from P. radiatus L..