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Construction and identification of human hepatocarcinoma cell cDNA library
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 2
- DOI:
- 10.3321/j.issn:1000-2790.2000.02.003
- Key Word:
- 人肝癌细胞;cDNA文库
Abstract： 目的构建人肝癌细胞cDNA表达文库.方法从人肝癌细胞中提取总RNA并纯化出mRNA,反转录合成第一、二链cDNA,除去小于500 bp cDNA片段,连接EcoRI人工接头,与去磷酸化的λgt11噬菌体左、右臂连接,体外包装后建成cDNA文库.取出一部分感染E.coli Y1090,测定文库大小并酶切鉴定插入cDNA片段的大小.结果构建成含1.2×106重组子人肝癌细胞cDNA文库,重组子平均插入外源片段长约1.5 kb.结论构建的文库合格,适合用于筛选目的cDNA克隆.
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