一种高效提取茄科类种子RNA的方法（英文）

Efficient Isolation of RNA from Seeds of Tomato and Chili Pepper

分子生物学是科学研究中最重要的科学领域,而RNA在分子生物学中又扮演着最重要的角色.因此,高效快速的RNA提取方法在分子生物学中成为一种关键技术.然而,茄科类植物（如番茄,辣椒）其种子中含有大量的蛋白质、多酚和多糖,并且易氧化,这些都严重影响RNA提取的质量.为了解决这些问题,本文以十二烷基肌氨酸钠/盐酸胍法为基础进行一定的改良,抽提的RNA的OD260/OD230均高于2.0,OD260/OD280在1.93-2.00之间,这表明RNA纯度很高,并且没有多酚多糖和蛋白质的污染.使用此方法抽提的RNA可应用于一系列的包括Northern印迹杂交和RT-PCR等的生物检测技术中.通过此方法能够从茄科植物的种子中提取出高产量、高质量的RNA,操作简单,成本较低.

RNA is an important object of molecular biology in scientific research. Extraction of high quality RNA in molecular biological method is a common user. High contents of protein,polyphenol and polysaccharide as well as the characteristics of easy oxidation make it a challenge for RNA extraction from seeds of Solanaceae plant,such as tomato and chili pepper. To solve these problems,a modified RNA extraction method is constructed based on saracosyl / guanidine hydrochloride method. In this report,the OD260/ OD230 ratio of extracted RNA is higher than 2. 0. Furthermore,the OD260/ OD280 ratios range from 1. 93-2. 00. This indicates that the RNA is of high purity and without polyphenol,polysaccharide or protein contamination. The extracted RNA could be used for a number of downstream analyses,including Northern blot hybridization,RT-PCR and so on. This method provides a method for isolation of high-quality and high-quantity RNA from seeds of Solanaceae plant. It is also a simple and low-cost extraction protocol.