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以长白山区野生膜荚黄芪根和叶为试材,参照TRIzoL说明书提取总RNA,采用SMART技术构建其全长cDNA文库,为分子水平上揭示膜荚黄芪次生代谢调控机制提供充足的基因资源。结果表明:原始文库滴度为4.8×105 cfu/mL,重组率接近99.1%,插入片段大于0.5kb,平均大小1.54kb。所构建的cDNA文库的库容量、克隆效率和全长率满足挖掘膜荚黄芪次生代谢相关基因的要求。
Based on the TRIzoL instructions, total RNA was extracted from the roots and leaves of wild A. membranaceus in Changbai Mountain. The full-length cDNA library was constructed by SMART technique, which provided abundant genetic resources for revealing the regulatory mechanism of the secondary metabolism of Radix Astragali. The results showed that the titer of the original library was 4.8 × 105 cfu / mL, the recombination rate was close to 99.1%, the inserted fragment was larger than 0.5kb and the average size was 1.54kb. The library capacity, cloning efficiency and full-length of the constructed cDNA library meet the requirements of excavating the genes related to secondary metabolism of Radix Astragali.