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Chinese Agricultural Science Bulletin ›› 2011, Vol. 27 ›› Issue (13): 126-129.

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An Efficient Method for Total RNA Extraction and LD_PCR Detection

  

  • Received:2010-11-11 Revised:2011-01-07 Online:2011-06-05 Published:2011-06-05

Abstract:

The aim was to extract RNA from Paris polyphylla var. yunnanensis and lay a foundation for studying the formation mechanism of Paris polyphylla var. yunnanensis. By modifying the method recommended by SDS/phenol for extracting total RNA from plant tissue rich in polysaccharidic and protein compounds, a simple and convenient method for extraction of total RNA from the embryo of Paris polyphylla var. yunnanensis containing abundant polysaccharides and protein was established. In RNA extraction buffer added to the steps of SDS was adjusted, combined with isopropanhol and sodium acetate at room temperature for more effective removal of precipitated polysaccharide, high concentrated SDS integrated with acid phenol/chloroform extraction were used to eliminate proteins; acetic acid-sodium acetate extraction buffer system to maintain a lower pH, inhibit the activity of RNA enzyme. The A260/A230 value of RNA extracted with imp roved method was all over 2.0 and the values of A260/A280 were between 1.8 and 2.0. The electrophoresis bands were cleared on agarosegel and integrity of RNA was good. The results showed that RNA obtained from the embryo of Paris polyphylla var. yunnanensis with this method had high purity and quality and could be used in molecular biological research, as LD-PCR and cDNA-library construction.