摘要
大鼠的肾脏组织经匀浆后,提取纯化总RNA和mRNA合成cDNA,进行甲基化,加人工接头,最后连入载体(λgt11)和外壳蛋白包装步骤制成肾脏组织的cDNA文库。插入载体的cDNA长度为0.5kb到8kb之间。经反复稀释,测定出该文库的效价为1.5×10~7pfu/mL。
Rat tissues were homogenized followed by purification of total RNAs and mRNAs, synthesizing cDNA, methylation, linking and finally ligation of the cDNA into λgt11 vector and packaging the length of the cDNA was 0.5-8kb. The titer was about 1.5×107pfu/mL.One positive cDNA clone was obtained by screening of the cDNA library using gluose transporter cDNA of rat liver as a probe. The fragments of the cDNA were obtained by digestion of the positive clone with E. coR1.
关键词
CDNA文库
CDNA克隆
分子杂交
筛选
cDNA library
cDNA clone
mRNA purification
Molecular hybridization and screening
