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[印迹杂交期间将DNA和RNA转移至硝酸纤维素滤膜上的一种新方法]

[A new method of DNA and RNA transfer onto nitrocellulose filters during blot hybridization].

作者信息

Likhtenshteĭn A V, Moiseev V L, Zaboĭkin M M

出版信息

Mol Biol (Mosk). 1987 Nov-Dec;21(6):1513-9.

PMID:2451780
Abstract

A quick (1-2 hour) method of DNA and RNA transfer onto nitrocellulose filters for subsequent blot-hybridization was elaborated. The main features of the method proposed are, firstly, almost complete exclusion of the mechanical impact on the gel and, secondly, addition to the transfer medium (20 X SSC) of a chaotropic agent, 0.5 M NaClO4. The latter results in a slight dissolution of the gel matrix and, on the other hand, somewhat increases the binding of the nucleic acid to the nitrocellulose. The method shortens significantly the time of DNA or RNA transfer at equal, or even higher, quality of hybridization.

摘要

阐述了一种将DNA和RNA快速(1 - 2小时)转移到硝酸纤维素滤膜上以便后续进行印迹杂交的方法。所提出方法的主要特点是,其一,几乎完全排除对凝胶的机械冲击;其二,在转移介质(20×SSC)中添加离液剂0.5M高氯酸钠。后者会导致凝胶基质略有溶解,另一方面,会在一定程度上增加核酸与硝酸纤维素的结合。该方法在杂交质量相同甚至更高的情况下,显著缩短了DNA或RNA的转移时间。

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