香花槐各组织总RNA提取方法的改良和优化

为探索香花槐各组织总RNA提取的最佳方法,以实验苗圃中的香花槐为实验材料,在比较了Trizol试剂法和常规CTAB法的基础上,建立了香花槐不同组织的最适提取方法.改良CTAB法利用高质量浓度的β-巯基乙醇和合适质量浓度的聚乙烯基吡咯烷酮(PVP)来防止RNA提取过程中多酚的氧化,然后增加苯酚/氯仿抽提次数来去除香花槐叶片和茎皮过多蛋白质.改良Trizol试剂法在根皮中加PVP研磨成白色粉末,可以有效地防止反应液褐化.各组织优化方法经琼脂糖凝胶电泳检测,所提取的总RNA的28S、18S条带清晰明亮,无降解;其A260nm/A280nm值为2.0,表明提取的总RNA质量较好.

Efficient improved methods for isolation of high quality total RNA from different tissues of Robinia pseudoacacia ’Idaho’

To search for the best methods,the different organs of Robinia pseudoacacia ’Idaho’ were used as materials to establish the improved methods for isolation of total RNA by comparing Trizol method and CTAB method.An improved CTAB method which can use the high concentration β-mercaptoethanol and 20 g/L PVP to eliminate the interferences of polysacchrides and the further phenol/chloroform extraction to eliminate the interferences of proteins of the leaf blade and stem of R.pseudoacacia ’Idaho’ was constructed in this study.The reaction solution could effectively prevent to brown by adding PVP in the root bark in the improved Trizol method.The optimized methods of each organ of R.pseudoacacia ’Idaho’ showed that 28S and 18S RNA bands were clearly visible and the A260 nm/A280 nm ratio of the isolated total RNA was 2.0,and which indicated that the total RNA quality was satisfying.