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多糖污染是从田间水稻叶片和未成熟种子等高含糖组织中提取高质量RNA的主要限制因素。我们介绍一种简单经济的RNA提取方法,它适合从水稻叶片和未成熟种子这些富含多糖的组织中提取总RNA。这种方法可以克服产率低和质量低的问题。RNA提取后经过电泳和分光光度计分析结果表明,每克水稻组织总RNA产率在520μg到800μg之间,OD260/230比值大于2.0,OD260/280比值大于1.9。提取的RNA可以用于体外反转录cDNA、mRNA分离和基因表达系列分析(serial analysis of gene expression,SAGE)库的构建。
Polysaccharide contamination is a major limiting factor for the extraction of high quality RNA from highly sugary tissues such as rice leaves and immature seeds in the field. We present a simple and economical RNA extraction method that is suitable for extracting total RNA from polysaccharide-rich tissues such as rice leaves and immature seeds. This method can overcome the problem of low yield and low quality. After RNA extraction, electrophoresis and spectrophotometer analysis showed that the total RNA yield per gram of rice was between 520μg and 800μg, the OD260 / 230 ratio was greater than 2.0, and the OD260 / 280 ratio was greater than 1.9. The extracted RNA can be used for in vitro reverse transcription of cDNA, mRNA isolation and serial analysis of gene expression (SAGE) library construction.