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. 1993 Oct;15(4):738-43.

Quantification of Specific mRNA by Flatbed Scintillation Counting of Dual-Labeled Dot Blots

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  • PMID: 8251177

Quantification of Specific mRNA by Flatbed Scintillation Counting of Dual-Labeled Dot Blots

C G Potter et al. Biotechniques. .

Abstract

A dual-labeling technique was developed for direct quantification of specific mRNA using a flatbed liquid scintillation counter. This method simultaneously measures cpm of 32P- and 35S-labeled probes bound to RNA dot blots and subtracts counts due to nonspecific background radioactivity bound to the filter. Probes for T-cell receptor and beta-actin (as the internal standard) were hybridized both separately and simultaneously to RNA isolated from five different sources. There was concordance between the radioactivity measured from single- and dual-hybridizations for each combination of 35S- and 32P-labeled probes. This methodology directly quantifies specific mRNA sequences bound to membranes and has potential for measuring gene dosage, without the need for re-probing or densitometric analysis.

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