广州管圆线虫成虫cDNA文库构建
Chinese journal of zoonoses（2003）
Abstract
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9.7×106 pfu/ml,重组效率达99.2%以上,文库扩增后滴度达9.13×109 pfu/ml.结论已成功构建了广州管圆线虫成虫cDNA文库.
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Key words
angiostrongylus cantonensis,adult worms,cdna library
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