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使用自制装置将DNA从凝胶电泳转移至滤膜。

Electrophoretic transfer of DNA from gels to filters by using a homemade apparatus.

作者信息

Pramatarova A, Michaud L, Gagnon F, Hamelin C

机构信息

Research and Virology Center, Armand-Frappier Institute, Laval-des-Rapides, Québec, Canada.

出版信息

Genet Anal Tech Appl. 1992 Feb;9(1):26-30. doi: 10.1016/1050-3862(92)90026-2.

Abstract

Electroblotting is a rapid and high-efficiency method of transferring DNA from gels to the variety of membranes available, in very short time frames when compared with capillary-blotting techniques and with much less preparation required. A simple apparatus made of two food savers, one dialysis membrane, one sponge, and two removable electrodes was used to transfer DNA molecules up to 21 kb, from agarose gels to nitrocellulose filters, in less than 30 min. A recombinant pAT153 plasmid carrying the human cytomegalovirus BamH I-P fragment (7.2 kb) was rapidly identified by hybridization, using our simplified transfer procedure.

摘要

电印迹是一种快速高效的方法,可在短时间内将DNA从凝胶转移到各种可用的膜上,与毛细管印迹技术相比,所需的准备工作要少得多。使用由两个食品保鲜袋、一个透析膜、一个海绵和两个可移动电极制成的简单装置,可在不到30分钟的时间内将长达21 kb的DNA分子从琼脂糖凝胶转移到硝酸纤维素滤膜上。通过杂交,利用我们简化的转移程序,快速鉴定出携带人巨细胞病毒BamH I - P片段(7.2 kb)的重组pAT153质粒。

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