通过与共价结合到纤维素上的过量互补DNA杂交对标记的珠蛋白信使RNA进行定量分析。

Quantitation of labeled globin messenger RNA by hybridization with excess complementary DNA covalently bound to cellulose.

作者信息

Levy S, Aviv H

出版信息

Biochemistry. 1976 May 4;15(9):1844-7. doi: 10.1021/bi00654a009.

Abstract

A method is described to quantitate labeled globin mRNA by hybridization with excess cDNA which was enzymatically polymerized on oligo(dT)-cellulose. In a large excess of cDNA-cellulose the rate of RNA hybridization was dependent on DNA concentration and not on RNA concentration. Nonhybridized RNA can be digested by RNase and washed from the cDNA which is covalently bound to cellulose. This enables the detection of labeled globin mRNA even when present in a porportion as low as 0.02-0.03% of the total RNA.

摘要

本文描述了一种通过与过量的、在寡聚（dT）-纤维素上酶促聚合的互补DNA（cDNA）杂交来定量标记珠蛋白信使核糖核酸（mRNA）的方法。在大量过量的cDNA-纤维素中，RNA杂交速率取决于DNA浓度而非RNA浓度。未杂交的RNA可被核糖核酸酶（RNase）消化，并从与纤维素共价结合的cDNA上洗脱下来。这使得即使标记珠蛋白mRNA在总RNA中的比例低至0.02 - 0.03%时也能被检测到。