舒峻, 李官成, 何小鹃, 李跃辉
Abstract
目的快速构建人鼻咽癌细胞株HNE2的定向cDNA文库.方法从人鼻咽癌细胞抽提总RNA磁珠法分离mRNA,以含SfiI酶切位点的oligo(dT)引物合成cDNA第一链,利用SMART(switching mechanism at 5'end of RNA transcript)技术,经LD-PCR合成双链cDNA,该物经SfiI酶切后分级分离,插入片段与λTripIEx2载体连接经体外包装成噬菌体cDNA文库.结果经鉴定,该文库含0.78×106个重组子,重组率＞96%;扩增文库滴度为1.02×109pfu/ml,插入片段平均长度为1.2kb.结论构建的人鼻咽癌细胞HEN2cDNA文库质量较好,有利于进一步筛选鼻咽癌特异性抗原基因.
A significance statement is not available in the OpenAlex record.
A contribution statement is not available in the OpenAlex record.
Method details are not available in the OpenAlex metadata.
Findings are not separately available in the OpenAlex metadata.
Limitations are not available in the OpenAlex metadata.
Application details are not available in the OpenAlex metadata.
目的快速构建人鼻咽癌细胞株HNE2的定向cDNA文库.方法从人鼻咽癌细胞抽提总RNA磁珠法分离mRNA,以含SfiI酶切位点的oligo(dT)引物合成cDNA第一链,利用SMART(switching mechanism at 5'end of RNA transcript)技术,经LD-PCR合成双链cDNA,该物经SfiI酶切后分级分离,插入片段与λTripIEx2载体连接经体外包装成噬菌体cDNA文库.结果经鉴定,该文库含0.78×106个重组子,重组率＞96%;扩增文库滴度为1.02×109pfu/ml,插入片段平均长度为1.2kb.结论构建的人鼻咽癌细胞HEN2cDNA文库质量较好,有利于进一步筛选鼻咽癌特异性抗原基因.
Key concepts: Nasopharyngeal carcinoma, Complementary DNA, cDNA library, Human cell, Computational biology, Biology, Gene, Medicine