Plant Diversity ›› 2008, Vol. 30 ›› Issue (05): 593-596.DOI: 10.3724 SP.J.1143.2008.07272

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An Effective Protocol for the Isolation of RNA from Cycad Leaves

LI Lu1 , FU Qian-Tang1 , 2 , YU Di-Qiu1   

  1. 1 Xishuangbanna Tropical Botanical Garden , Chinese Academy of Sciences, Kunming 650223 , China;
    2 Graduate University of Chinese Academy of Sciences, Beijing 100049 , China
  • Received:2007-11-21 Online:2008-10-25 Published:2008-10-25

Abstract: Conventional RNA extraction methods have been shown to produce poor-quality RNA when applied to Cycas revoluta because of abundant secondary metabolites included. With modification of the standard cetyltrimethylammonium bromide (CTAB) method by adding borax (disodium tetraborate decahydrate ) andβ-mercaptoethanol to eliminate the interference of polyphenol and polysaccharides , an effective protocol was developed . This modified protocol could extract high quantities of RNA from cycad leaves at about 930μg per gram of fresh weight . Both A260􊄯230 and A260􊄯280 ratios were around 2, indicating that RNA is of high quality. The RNA quality was confirmed by Northern blotting analysis and reverse transcription polymerase chain reaction (RT-PCR). The modified protocol could be successfully extended to other plants containing secondary metabolites .

Key words: Cycas revoluta

CLC Number: