EXTRACTION OF HIGH-QUALITY RNA AND CONSTRUCTION OF A CDNA LIBRARY FROM FRUITS OF LYCIUM BARBARUM LINNAEUS (FRUCTUS LYCII)

ABSTRACTHigh-quality RNA isolation is the prerequisite for molecular analysis. In the present study, we used the RNAiso™ for Polysaccharide-rich Plant Tissue solution to extract high-quality RNA from fruits of Lycium barbarum Linnaeus (Fructus Lycii), a popular traditional Chinese herbal medicine used for centuries. The RNA was illustrated to be intact, without protein contamination. Based on the RNA isolated, the gene encoding β-carotene hydroxylase (BCH) was partially cloned by performing reverse transcription-polymerase chain reaction (RT-PCR). In addition, a high quality cDNA library with 6.0×106 clones in total was successfully constructed. The results indicated that this method could be reliably used in isolating high-purity RNA from Fructus Lycii. The outcome will be helpful to the studies of carotenoids or biosynthesis other pharmaceutical elements in Fructus Lycii.