红花花瓣总RNA提取方法的比较研究

Study on Total RNA Extraction from Safflower Petals

红花花瓣富含多糖、多酚和花色素苷等次生代谢产物,严重影响花瓣总RNA的提取。为探索适合红花花瓣总RNA的提取方法,以4种红花不同时期的花瓣为材料,通过对4种RNA提取方法,即RNA提取试剂盒、RNAiso Plus试剂法、改良的CTAB法和SDS法进行比较研究。结果表明:RNAiso Plus试剂法效果最好,提取的总RNA完整性好,纯度高、浓度高。通过RT-PCR验证,所提取的RNA达到基因克隆和实时定量PCR等分子生物学实验要求。

It is difficult to isolate total RNA from safflower petals because of a great deal of polysaccharides , polyphenolics ,anthocyanins and other metabolites .To explore suitable RNA extraction method of safflower petals ,total RNA was isolated with the method of improved CTAB method ,RNA extraction kit ,RNAiso Plus Reagent and SDS method .The results showed that RNA extracted by total RNAiso Plus reagent method had best integrity ,high purity ,high concentrations .The total RNA isolated by this method was proved to be rela‐tively intact and high purity .The isolated RNA could be used in quantitative real‐time PCR and other molecular biology experiments by RT‐PCR analysis .