Extraction method of RNA from Pinus tabulaeformis Carr mature embryo | 油松成熟胚总RNA的提取方法__
2010
Zhao Baihui, Beijing Forestry University, Beijing (China) , College of Biological Sciences and Biotechnology | Zheng Caixia, Beijing Forestry University, Beijing (China) , College of Biological Sciences and Biotechnology | Xu Lili, Beijing Forestry University, Beijing (China) , College of Biological Sciences and Biotechnology
Китайский. [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,_A__260_/A__280_为1.97,A__260_/A__230_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。
Показать больше [+] Меньше [-]Английский. [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,_A__260_/A__280_ _was 1.97 and _A__260_/A___230_ 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._
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