摘要
采用4种方法分别对小麦幼叶、幼茎、幼根及乳熟期籽粒的总RNA进行了提取和分析.结果表明,4种方法所提取的总RNA均能满足一般的下游操作,其中TRIzol法提取的总RNA产量最高,用该法提取的小麦叶片总RNA产量为其他方法的2～4倍;天为时代试剂盒提取时间短,可在常温下操作;而醋酸钠/乙醇沉淀法具有提取步骤少、操作时间短、可用于大量样品提取且价格便宜等优点,适于小麦不同器官的总RNA提取;经紫外分光光度计检测,各样品总RNA的A260/A280均在1.88～2.09,表明所提取的总RNA中没有蛋白质及其他有机溶剂的污染,且A260/A230在1.89～2.41,表明盐类小分子的污染也较少,所提取的总RNA质量较高.除了细胞质rRNA外,小麦中还存在叶绿体rRNA,且叶绿体rRNA占总rRNA的比例较高,在甲醛变性琼脂糖凝胶电泳时,幼叶、幼茎比幼根明显多出2条带.
Total RNA was extracted from young leaf,young stem,radicle,and immature se by four different methods respectively. The results showed that all of the purified total RNA standard and general downstream applications through purity and quantity analysis. Among ods,TRhol could get the highest production of total RNA,which was as much as 2-4 times t methods. Tianweishidai kit needed less time, and could operate at room temperature. Due to f ed W of ere wheat up to these methhat of other ewer steps, less time,lower price and suitable for large quantity samples,NaAC/ethanol was a desirable method to extract total RNA from different organs in wheat. Detected by UhravioIet spectrophotometer,A260/A280 was 1.88-2.09 and A260/A230 was 1.89-2.41 ,which showed the purity of total RNA was better. Analyzed by formaldehyde denaturalized gel eIectrophoresis,totaI wheat RNA contained large percentage of chloroplast rRNA other than cytoplasm rRNA,which resuhed in two more rRNA bands in young leaf and young stem than those in radicle.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2005年第10期29-32,共4页
Journal of Northwest A&F University(Natural Science Edition)
基金
教育部高校科技创新工程重大项目培养资金
关键词
小麦
总RNA含量
提取方法
wheat
total RNA content
extraction methods
