广州管圆线虫成虫cDNA文库构建

目的　构建广州管圆线虫成虫cDNA文库。方法　提取广州管圆线虫成虫总RNA ,用Clontech公司SMARTTMcDNA文库构建试剂盒反转录合成第一链cDNA ,然后通过长距离PCR方法合成双链cDNA并扩增 ;PCR产物与λTripIE× 2载体连接并包装 ,建成未扩增文库 ;检测未扩增文库滴度和重组效率后 ,进行文库扩增及测定扩增文库的滴度。结果　经测定 ,未扩增文库滴度为 9 7× 1 0 6 pfu/ml,重组效率达 99 2 %以上 ,文库扩增后滴度达 9 1 3× 1 0 9pfu/ml。结论　已成功构建了广州管圆线虫成虫cDNA文库

Construction of a cDNA library from adult worms of Angiostrongylus cantonensis

Aim To construct a cDNA library from adult worms of Angiostronglus cantonensis Methods The total RNA from the adult worms of Angiostronglus cantonensis was extracted After that we retro transcribed to synthesize the single strand cDNA using SMART oligo,then synthesized and amplified double-strand cDNA by long-distance PCR (LD-PCR) The PCR products were ligated intoλTripIEx2 vector The unamplified library wasn't completed until the recombinants were packaged by packaging extracts The capacity and the recombination rate about the unamplified library were detected and then amplified it Results The unamplified library capacity was 9 7×10 6 pfu/ml,the recombination rate was more than 99 2% After the library was amplified,its capacity was 9 13×10 9 pfu/ml Conclusion The cDNA library of adult A cantonensis has been constructed using SMART LD-PCR technology in this experiment for the first time