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用于杂交实验的双链DNA电转印:在脉冲场凝胶电泳后10分钟内DNA转移完成。

Electroblotting of double-stranded DNA for hybridization experiments: DNA transfer is complete within 10 minutes after pulsed-field gel electrophoresis.

作者信息

Ishihara H, Shikita M

机构信息

Division of Chemical Pharmacology, National Institute of Radiological Science, Chiba, Japan.

出版信息

Anal Biochem. 1990 Feb 1;184(2):207-12. doi: 10.1016/0003-2697(90)90670-5.

Abstract

A rapid, simple, and efficient method for DNA blotting is presented. This method is characteristic in that DNA is transferred without denaturation from an electrophoretic gel to a membrane in low-salt concentrations and denatured on the membrane after blotting. More than 89% of double-stranded DNAs ranging in size from 75 to 1.9 million bp can simultaneously be transferred from the gel to a positively charged nylon membrane within 10 min. The present "low-salt electroblotting" method is superior to other blotting methods in that it saves time and labor and its high and even transfer efficiency makes it useful for hybridization analysis.

摘要

本文介绍了一种快速、简单且高效的DNA印迹法。该方法的特点是DNA在低盐浓度下从电泳凝胶转移至膜上时无需变性,而是在印迹后在膜上进行变性。大小在75至190万碱基对之间的双链DNA中,超过89%能够在10分钟内同时从凝胶转移至带正电荷的尼龙膜上。目前的“低盐电印迹”方法在节省时间和人力方面优于其他印迹方法,其高且均匀的转移效率使其适用于杂交分析。

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