CAJ | 학술논문

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520 μg到800 μg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.
다당오염시종전간수도협편화미성숙충자등고함당조직중제취고질량RNA적주요한제인소.아문개소일충간단경제적RNA제취방법,타괄합종수도협편화미성숙충자저사부함다당적조직중제취총RNA.저충방법가이극복산솔저화질량저적문제.RNA제취후경과전영화분광광도계분석결과표명,매극수도조직총RNA산솔재520 μg도800 μg지간,OD260/230비치대우2.0,OD260/280비치대우1.9.제취적RNA가이용우체외반전록cDNA、mRNA분리화기인표체계렬분석(serial analysis of gene expression,SAGE)고적구건.
Polysaccharides contamination is the major limitative factor for high-quality RNA extraction from the high level polysaccharides tissues in field-grown rice leaves and immature seeds. Here we introduce a simple and inexpensive method for total RNA extraction. It is suitable for extracting total RNA from rice leaves and immature seeds rich in polysaccharides . This method overcame the shortcoming of low yield and low quality . After extraction, the RNA was analyzed by spectrophotometric readings and electrophoresis in denaturing agarose gels. The result indicates that total RNA yield is ranging from 520 μg to 800 μg per gram of tissue with an averaged purity measured as OD260/230 of more than 2.00 and OD260/280 more than 1.900. The extracted RNA can be used in exclusive amplification to cDNA template RT-PCR, mRNA isolation and SAGE (serial analysis of gene expression) library construction.