Comparison of the methods of RNA isolation from <italic>Juglans regia</italic> L. buds

doi:10.1007/s11703-011-1074-6

Front Agric Chin

2011, Vol. 5

Issue (2) : 221-224 https://doi.org/10.1007/s11703-011-1074-6

RESEARCH ARTICLE

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

Fuqiang HE, Hongxia WANG, Xiaobo SONG, Zhihua ZHANG(

)

Mountainous Areas Research Institute, Hebei Agricultural University, Baoding 071001, China

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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.

Keywords Juglans regia L. buds total RNA RP3301 kit

Corresponding Author(s): ZHANG Zhihua,Email:shylg@hebau.edu.cn

Issue Date: 05 June 2011

Cite this article:

Fuqiang HE,Hongxia WANG,Xiaobo SONG, et al. Comparison of the methods of RNA isolation from Juglans regia L. buds[J]. Front Agric Chin, 2011, 5(2): 221-224.

URL:

https://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1074-6
https://academic.hep.com.cn/fag/EN/Y2011/V5/I2/221

Fig.1 The agarose gel electrophoresis for RNA extracted by different methods. Note: 1-4 are in CTAB method (1-2 in 1% CTAB, 3-4 in 2% CTAB). 5-6 are in TRIzol method. 7-8 are in D326A reagent. 9-10 are in RP3301 kit. 1, 3, 5, 7 and 9 stand for Zhonglin. 5, 2, 4, 6, 8 and 10 stand for Qingxiang.

Tab.1 OD, OD and yield of RNA extracted by different methods

Fig.2 The RT-PCR bands of No.9 and No.10 RNA. Note: M stands for marker DL2000, Lanes 1 and 3 mean RT-PCR without M-MLV, and Lanes 2 and 4 mean RT-PCR with M-MLV. 1-2 stand for Zhonglin 5 and 3-4 stand for Xiangling 5.

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