申请人:INSTITUT PASTEUR DE LILLE
公开号:EP0882801A1
公开(公告)日:1998-12-09
This invention consists in a method for selecting cDNA fragments generated by differential display or RNA fingerprinting comprising the steps of :
a) excising a differential band of the migration gel used for differential display or RNA fingerprinting and amplifying the corresponding cDNA fragments by PCR,
b) submitting the PCR amplified cDNA fragments to gel electrophoresis in the presence of at least one polymer-coupled DNA binding agent and/or a polypeptide capable of DNA binding and mobility retardation,
c) visualizing resolved cDNA fragments by means of a staining agent or radioactive labeling,
d) excising the desired bands separated in step b) and directly amplifying the corresponding cDNA fragments by PCR,
e) optionally purifying and analysing the amplified PCR products.
本发明包括一种选择通过差异显示或 RNA 指纹法生成的 cDNA 片段的方法,包括以下步骤 :
a) 从用于差异显示或 RNA 指纹的迁移凝胶中切除差异带,并通过 PCR 扩增相应的 cDNA 片段、
b) 将 PCR 扩增的 cDNA 片段在至少一种聚合偶联 DNA 结合剂和/或具有 DNA 结合和迁移率延迟能力的多肽存在下进行凝胶电泳、
c) 通过染色剂或放射性标记对已分离的 cDNA 片段进行可视化、
d) 去除在步骤 b) 中分离出的所需条带,并通过 PCR 直接扩增相应的 cDNA 片段、
e) 对扩增的 PCR 产物进行纯化和分析。