mCTAB-dLiCl method for efficiently extracting RNA from various tissues of cultivated peanuts and its vertification

【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.