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| A Novel Method of Total RNA Extraction from Leaves of Cinnamomum longepaniculatum |
| LIN Na1,2, GAO Ji-hai1, AI Tao-bo1, FAN Lin-hong1, WANG Sheng-hua1, CHEN Fang1 |
1. Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China;
2. Institute of Biology, Yibin University, Yibin 644000, China |
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Abstract The leaves of Cinnamomum longepaniculatum are rich in lipids, polyphenols and polysaccharides. Various conventional lysis reagent (Trizol reagents, bentonite, CTAB and Tris-SDS- guanidinium isothiocyanate) methods were tested for extraction of the total Cinnamomum longepaniculatum RNA without ideal results. Based on characters of the leaves containing multiple secondary metabolites to inference RNA purification, a novel modified method was tested: the pretreatment liquid and ethyl acetate were used firstly to reduce lysate fluid viscosity; chloroform was replaced by dichloromethane in RNA extraction process to eliminate effect of hydrophobic impurity; then high concentration of mixed salt were used to take off sugars. Results showed that the new modified method of total RNA extraction from Cinnamomum longepaniculatum leaves was purified, with no obvious degrade, the A260/A280 and A260/A230 were about 2.0, productivity was 580 μg /g. The RNA from the novel method was amplified through RT-PCR and with a 113bp specific sequence. The data identified RNA integrity, purity and production efficiency were improved by the novel modified method, and this experiment has determined a new modified method for extraction of total RNA from Cinnamomum longepaniculatum leaves.
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Received: 17 December 2012
Published: 25 May 2013
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