摘要
用异硫氰酸胍法从孢子化 7h的卵囊中提取总 RNA,链合成引物用置换合成法合成总 c DNA,c DNA与 E再用 oligo( d T) n-纤维素从总 RNA中分离 m RNA。m RNA以 Not -olig( d T) 15作第一链合成引物用置换合成法合成总 c DNA,c DNA与 Eco R Adaptors的连接后 ,经 Not 酶切 ,再与 Eco R 和 Not 预切好的 λ载体连接 ,构建成鸡球虫 c DNA文库。
Total RNA of Eimeria tenella was extracted by the guanikine thiocynate procedure from sporulating-7 h oocysts,and mRNA was isolated with oligo(dT) n-cellulose.First strand cDNA synthesis was driven by AMV from mRNA USING OLIGO(Dt) 15 primers containing Not Ⅰ restriction sites at their 5' ends,and second strand replacement synthesis was followed directly.After addition of EcoR Ⅰ adaptors to cDNA molecules and digestio wkth Not Ⅰ restriction enzyme,large-sized cDNA was then cloned into λgt Ⅱ S-Not Vector.It was showed that the cDNA library is a partial libaray.
出处
《动物医学进展》
CSCD
2002年第5期64-65,共2页
Progress In Veterinary Medicine
基金
国家自然科学基金 (30 170 6 95 )
