Comparison of the methods of RNA isolation from Juglans regia L. buds

Abstract

In this paper, four RNA extraction methods (CTAB method, TRIzol method, D326A reagent and RP3301 Kit) for walnut buds were compared by agarose gel electrophoresis and spectrophotometry, and the optimal method was verified further by Reverse transcription and PCR amplification (RT-PCR). There was a certain degree of degradation for the extracted RNA by CTAB method, TRIzol method and D326A reagent. Moreover, protein was found in the CTAB method. For the three unsatisfactory methods, longer operation time could be the main reason that RNA could not be separated in time with polyphenols, polysaccharides and other secondary metabolites. RP3301 kit was suitable for the RNA extraction from walnut buds with high purity, simplicity, innocuity and no degradation. This research will provide a fundamental basis for the cloning and expression analysis of genes and other follow-up tests.

Key words： Juglans regia L.; buds; total RNA; RP3301 kit

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