改良热硼酸法高效提取苹果果实RNAEffective extraction of total RNA in apple flesh with improved hot borate method
姚玉新,赵玲玲,郝玉金,翟衡
摘要(Abstract)：
苹果果实、尤其是成熟期的果实中多糖和其他次生物质含量高,难以提取RNA。对此,在原热硼酸法的基础上进行了改进,通过添加提取辅助剂并根据辅助剂特性调整提取步骤,高效、稳定地从苹果果实、尤其是后期果实中提取到了高质量的RNA。所得RNA的A260/A230大于2.0,A260/A280为1.8-2.1,并且RNA产量高,其中前期果实RNA产率在13.40μg/g以上,后期果实RNA产率在7.02μg/g以上,完全可以满足RT-PCR、cDNA-AFLP等分子生物学实验的要求。
关键词(KeyWords)： 苹果果实;总RNA;改良热硼酸法
基金项目(Foundation): 山东省2003良种产业化项目(编号:32271)。
作者(Author): 姚玉新,赵玲玲,郝玉金,翟衡
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DOI:
参考文献(References)：
- [1]M anning K.Isolation ofnucleicacidsfrom plantsbydifferentialsol鄄ventprecipitation[J].AnalBiochem,1991,195:45-50.
- [2]H unterDA,Reid M S.A Simple and rapid m ethod forisolating high qualityRNA from flowerpetals[J].Acta H ort,2002,319:543.
- [3]W an CY,W ilkins TA.A M odified H otBorate M ethod Significantly Enhances the Yield ofH igh-Quality RNA from Cotton(Gossypium hirsutum L.)[J].AnalyticalBiocem,1994,223:7-12.
- [4]蒋建雄,张天真.利用CTAB/酸酚法提取棉花组织总RNA[J].棉花学报,2003,15(3):166-167.
- [5]Ainsworth C.Isolation ofRNA from floral tissue ofRumex aetosa(Sorrel)[J].PlantM olBiolReptr,1994,12:198-203.
- [6]Chen FS,Brown SK,W eeden NF.A DNA extraction protocolfrom varioustissuesin woody species[J].JPlantCrops,2000,28(1):30-34.
- [7]Loom is W D.Overcoming problem sofphenolicsand quinines in the isolation ofplantenzym es and organelles[J].M eth Enzym ol,1974,31:528-545.
- [8]BahloulM,Burkard G.An im proved m ethod forthe isolation oftotal RNA from spruce tissues[J].PlantM olBiolReptr,1993,11:211-215.