熊发前1 2 * 刘俊仙3 * 王丛丛4 贺梁琼1 韩柱强1 蒋 菁1 钟瑞春1
李 忠1 唐秀梅1 唐荣华1 2 * *
|单位： （1 广西农业科学院 经济作物研究所 南宁 530007；2 广西作物遗传改良生物技术重点开放实验室 南宁 530007；3 广西农业科学院 甘蔗研究所 南宁 530007；4 仲恺农业工程学院 广州 510225）|
|关键词： 花生 RNA提取 mCTAB-dLiCl法 组织部位|
|出版年,卷(期):页码：2013 ,44 ( 11 )： 页码：1781-1784|
【Objective】RNA extraction methods for different tissue of peanut were explored to provide references for peanut molecular biology research. 【Method】An improved mCTAB-dLiCl method was designed to extract RNA on the basis of published CTAB method. The mCTAB-dLiCl method used high concentrations of KAC to remove polysaccharide from tissues and used LiCl twice to precipitate RNA. The quality and purity of RNA were checked by agarose gel electrophoresis and nucleic acid protein analyzer， respectively. The final quality of RNA was confirmed by reverse transcription， RT-PCR and cDNA-SCoT analysis. 【Result】Using the mCTAB-dLiCl method， the yield of RNA and the ratio of A260/A280 of RNA were 554.80 μg/g and 1.89， respectively. No visible DNA contamination was observed through the non-denaturing agarose-gel electrophoresis. The two bands having the ratio of 2∶1 corresponding to 28S and 18S were observed. Also no smear was observed in bands which showed no polysaccharide contamination in RNA. Through testifing by reverse transcription， RT-PCR and cDNA-SCoT analysis， the extraeting RNA achieved good amplification effects， and the obtained RNA and cDNA exhibited good quality. 【Conclusion】mCTAB-dLiCl RNA extraction method， a highly efficient method for extracting RNA from various peanut tissues， could be applied to peanut molecular biology research.