一种无需DEPC提取高质量棉花总RNA的方法

A Good Method without DEPC for Extraction of High-quality Total RNA from Cotton

DEPC做为RNAnase抑制剂常用于RNA的提取，但其本身具有毒性，容易对操作者产生伤害并易造成环境污染。该实验以改良CTAB法为基础，提高β-巯基乙醇的浓度到10％，无需DEPC就能提取高质量的RNA。分别以不同陆地棉为材料提取了棉花的总RNA，测得OD260／OD：80在1．7—2．1之间，且大部分样品都能清晰看到3条RNA条带，将此RNA反转录获得的cDNA为模板克隆到了棉花ATP合成酶β亚基基因，说明此方法提取的棉花总RNA既能满足分子生物学实验要求又大大地减少了毒害作用。

DEPC as RNAnase inhibitor is often used in the extraction of RNA, but it is toxic to the operator and easily causes environmental pollution. In order to improve the CTAB -based method and enhance β-mereaptoethanol concentration to 10%, the author extracted high -quality RNA without DEPC. In this experiment, the total RNAs were respectively extracted from various upland cotton varieties, their OD260/OD280 ranged from 1.7 to 2.1, and three RNA bands could be clearly seen in most samples, the cDNA from the reverse transcription of this RNA have been cloned to cotton β-subunit gene of ATP synthase. This method for the extraction of total RNA from cotton is able to meet the requirement of molecular biology test and greatly reduce the toxicity of DEPC.