摘要
以宁夏枸杞叶片为材料,采用异硫氰酸胍—酚—氯仿法提取完整的总RNA,利用PolyATractmRNAIsolationSystem分离mRNA,然后用UniversalRibocloneRcDNASynthesisSystem合成双链cDNA,在cDNA末端连接上EcoRⅠ接头,并与λExCell载体连接,利用PackageneLambdaDNAPackagingSystem进行包装,在国内外首次构建出滴度为2 78×105pfu/mL,重组效率为88 1%的枸杞叶片cDNA文库。
<Abstrcat> Total RNA of leaf of Lycium barbarum L. was isolated using guanidinium isothioccyanate and phenol-chloroform. mRNA was prepared by PolyATract mRNA Isolation Systems (Promega). Full-length cDNA was synthesized by Universal RiboClone cDNA Synthesis System(Promega) and then cloned into phage λExcell vector. After Pachage in Packagene Lambda DNA Packaging System, the capacities of the libraries were measured. The capacity of the library was 2.78×10~5 pfu/ml. The recombination rate reached 88.1%.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2005年第2期155-157,161,共4页
Journal of Jilin Agricultural University
基金
吉林省科技厅资助项目(20020641)。
关键词
枸杞
叶片
CDNA文库
Lycium barbarum L.
leaf
cDNA library
