甜高粱茎秆总RNA提取方法的比较
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摘要
甜高粱是最具潜力的能源作物之一,新鲜的甜高粱茎秆中含有大量汁液且富含多糖和多酚成分,很难获得高质量的总RNA。以甜高粱Rio的茎秆为实验材料,利用Trizol法、SDS法、改良CTAB法和RNAiso法分别提取新鲜茎秆总RNA,比较几种方法获取的总RNA质量与纯度。实验结果证明,RNAiso改良法提取的茎秆总RNA质量高,完整性好,总RNA产物可直接用于建库和转录组测序等后续实验。
Sweet sorghum is a kind of promising energy crops.There are large amounts of juice andrich in polysaccharides and polyphenols in fresh stem of sweet sorghum,so it is very difficult to obtain high quality total RNA by common RNA extraction methods from the fresh stem.In this study,the total RNA of Rio stem was extracted by Trizol method,SDS method,modified CTAB method and RNAiso method respectively.The results showed that the total RNA extracted usingRNAiso method is of high quality and good integrity,and can be used directly for subsequent experiments such as database construction and transcriptome sequencing.
Sweet sorghum is a kind of promising energy crops.There are large amounts of juice andrich in polysaccharides and polyphenols in fresh stem of sweet sorghum,so it is very difficult to obtain high quality total RNA by common RNA extraction methods from the fresh stem.In this study,the total RNA of Rio stem was extracted by Trizol method,SDS method,modified CTAB method and RNAiso method respectively.The results showed that the total RNA extracted usingRNAiso method is of high quality and good integrity,and can be used directly for subsequent experiments such as database construction and transcriptome sequencing.
引文
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[3]王爱勤,范业赓,赵晓艳,等.甘蔗茎RNA提取方法的比较[J].广西农业生物科学,2007,262(2):175-178.
[4]魏鑫,李玥莹,关尔鑫.甜高粱总RNA提取方法的比较研究[J].江苏农业科学,2011,39(3):45-47.
[5]尹慧,陈莉,李晓艳,等.百合叶片总RNA提取方法比较及优化[J].中国农业大学学报,2008,13(4):41-45.
[6]Mueller O,Hahnenberger K,Dittmann M,et al.A microfluidic system for high-speed reproducible DNA sizing and quantitation[J].Electrophoresis,2000,21(1):128~34.
[7]Lightfoot S.Quantitation comparison of total RNA using the Agilent 2100 bioanalyzer,ribogreenanalysis and UV spectrometry[M].Agilent Application Note,2002,Publication number:5988~7650EN.
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[9]袁明珠,温柔,刘吉升,等.几种植物材料中总RNA的提取[J].分子植物育种,2005(3):285~292.
[10]王英,羊玉花,邱海燕,等.甘蔗种质总RNA提取方法的研究[J].中国农学通报,2009,25(24):68~72.