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玉米成熟子粒RNA提取方法的改良与优化

High Quality RNA Isolation from Mature Seeds of Maize Based on Modified and Optimized Protocols
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摘要 玉米成熟子粒含有较多的多糖、蛋白质和脂类,传统的RNA提取方法难以提取到高质量的RNA。以授粉后28 d灌浆后期的玉米子粒为试材,以授粉后7 d的玉米子粒为对照,对目前国内外常用的Trizol法、Biozol法、RNAiso Plus法、三种硅胶柱商业试剂盒总RNA提取方法以及本研究建立的改良提取方法进行比较。结果表明,以授粉后7 d的玉米子粒为材料时,各种方法都能获得较好提取效果;以灌浆后期接近成熟玉米子粒为试材时,采用本研究改良或新建立的提取法,可以大幅降低提取成本,快速提取高质量和高得率的RNA。 It is usually difficult to isolate high-quality RNA from mature seeds of maize(Zea mays L) using tra- ditional methods due to the abundance of polysaccharides, protein and lipid. Mature maize seeds of 28 clays after pollination were used for RNA isolation and early developing seeds of 7 days after pollination were used as the con- trol. The methods of Trizol, Biozol, RNAiso Plus and three silica columns based commercial RN A isolation kits were compared for RNA isolation with their modified versions and the new optimized protocol established. The results showed that while high-quality RNA was isolated from developing seeds of 7 days after pollination using all the test- ed methods, only the modified protocols and our new protocol resulted in high-quality and high-yieht RNA from ma- lure maize seeds of 28 DAP. In addition, the newly developed protocol was also much more cost effective and time saving compared to all other tested and modified methods.
作者 张园园 赵吉强 姜丽君 史艳慧 陈磊 郭善利 宋建成 Paula E Jameson ZHANG Yuan-yuan, ZHAO Ji-qiang, JIANG Li-jun, SHI Yah-hui, CHEN Lei, GUO Shan-li, SONG Jian-cheng, Paula E Jameson (1. School of Life Sciences, Yantai University, Yantai 264005, China; 2. School of Biological Sciences, University of Canterbury, Christchurch 8140, New Zealand)
出处 《玉米科学》 CAS CSCD 北大核心 2015年第4期38-42,47共6页 Journal of Maize Sciences
基金 国家自然科学基金(3137616)、山东省自然科学基金(ZR2011CM044,ZR2011CM006)
关键词 玉米 子粒 RNA提取 Maize Seed RNA isolation
作者简介 张园园(1984-),女,河南安阳人,硕士,从事植物分子发育生物学研究。Tel:13791292205E-mail:zyy1984126@163.com. 宋建成为本文通讯作者。Email:jcsong88@yahoo.com
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