蛋白质对用Hoechst 33258测定DNA的影响。

The protein effect on determination of DNA with Hoechst 33258.

作者信息

Moe D, Garbarsch C, Kirkeby S

机构信息

Department of Oral Function, Dental School, Faculty of Health Sciences, University of Copenhagen, Denmark.

出版信息

J Biochem Biophys Methods. 1994 Jun;28(4):263-76. doi: 10.1016/0165-022x(94)90002-7.

DOI:10.1016/0165-022x(94)90002-7

PMID:7525683

Abstract

The present study was designed to afford a critical review of the effect of proteins on the Hoechst 33258 method for determination of DNA of crude homogenates. A considerable effect of proteins on the fluorescence was observed when the concentration exceeded 100 micrograms BSA equivalent protein. Below that value, practically no effect of proteins was noted. We used proteinase K to remove the proteins, but dilution of homogenates could be used as well. Moreover, we found that the concentration of the fluorochrome should be between 1 microgram and 2 micrograms when microgram levels DNA are to be determined.

摘要

本研究旨在对蛋白质对用Hoechst 33258法测定粗匀浆DNA的影响进行批判性综述。当蛋白质浓度超过100微克牛血清白蛋白（BSA）当量蛋白时，观察到蛋白质对荧光有相当大的影响。低于该值时，实际上未注意到蛋白质有影响。我们使用蛋白酶K去除蛋白质，但也可以使用匀浆稀释的方法。此外，我们发现，当要测定微克水平的DNA时，荧光染料的浓度应在1微克至2微克之间。