油松成熟胚总RNA的提取方法  

Extraction Method of RNA from Pinus tabulaeformis Carr Mature Embryo

作　　者：赵百慧[1] 郑彩霞[1] 徐利利[1] 赵楠[1] 徐桂娟[1] 

机构地区：[1]北京林业大学生物科学与技术学院,北京100083

出　　处：《安徽农业科学》2010年第6期2812-2814,共3页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(30671655)

摘　　要：[目的]对油松成熟胚总RNA的提取方法进行比较,以期得到完整的高质量油松成熟胚RNA。[方法]以油松成熟胚为材料,用SDS法、Trizol法和CTAB法及改良的SDS法对油松成熟胚总RNA进行提取,以紫外分光光度计测OD值,并用琼脂糖凝胶电泳检验提取效果。[结果]改良SDS法提取的RNA产率高,完整性好,A260/A280为1.97,A260/A230为2.08,28 S、18 S、5 S条带清晰且28 S亮度是18 S的2倍。通过RT-PCR检测,ds cDNA片段的弥散带分布范围在0.3～3.0 kb,进一步验证改良的SDS法可获得高质量的RNA,用于后续的分子生物学研究。[结论]改良SDS法成本低,操作简单,提取时间短,RNA质量好,是多酚多糖植物总RNA提取比较有效的方法。[Objective]The research aimed to compare the extraction method of total RNA from Pinus tabulaeformis Carr,and to obtain high quality total RNA of P.tabulaeformis Carr mature embryo.[Method]SDS mehtod,CTAB method,Trizol method and improved SDS mehtod were employed to isolate total RNA from P.tabulaeformis Carr mature embryo.And the qaulity of RNA was examined with OD value and agarose gel electrophoresis.[Result]The purity of RNA extracted was high by improved SDS mehtod,A260/A280 was 1.97 and A260/A230 was 2.08.The bands of 28 S,18 S and 5 S were clear and the brightness of 28 S bands was twice of the bands of 18 S.After RT-PCR,ds cDNA bands size was range from 0.3 to 3.0 kb,the results further demonstrated that the purity and integrality of total RNA isolated using improved SDS method were significantly satisfactory for the demands of molecular biological research.[Conclusion]Improved SDS mehtod had advantages of low-cost,simple-operation,short-time,good quality of extracted RNA,and it was an effective method of the extraction of total RNA of polysaccharide and polyphenol plants.

关 键 词：油松 成熟胚 RNA 改良SDS法 

分 类 号：S791.254[农业科学—林木遗传育种]