赤子爱胜蚓全长cDNA文库的构建及初步鉴定

Construction and primary identification of full-length cDNA library of Eisenia foetida

以赤子爱胜蚓(Eisenia foetida)为材料,λTriplex2TM为载体,利用SMART技术构建全长cDNA文库,从而筛选全长目的基因并研究其结构和功能。经测定原始文库滴度为1.3×106pfu/mL,扩增总文库滴度为1.0×108pfu/mL,重组率达到99%以上,插入片段长度在1.1 kb左右。该文库的建立为功能性新基因的发现奠定了坚实的基础。

To obtain a full - length cDNA library of Eisenia foetida for the purpose of gene screening and functioning study, total RNA extraction method from Eisenia foetida used is TRIZOL Reagent to construct a cDNA library by the carrier of λTriplex2^TM using the SMART cDNA Library Construction Kit. The quality of the constructed full - length cDNA library was evaluated. The primary library showed a titer of 1.3×10^6 pfu/mL of the cDNA insets, and the amplified library has a titer of 1.0 × 10^8. The recombinant rates were about 99 % and the average length of the cDNA inserts was about 1.1 kb. This constructed cDNA full - length library provided an essential resource for the further gene functional studies and gene discoveries of Eisenia foetida.