A simple subtractive hybridization technique employing photoactivatable biotin and phenol extraction

Abstract

We report a simple method for nucleic acid subtractive hybridization, that involves hybridization of untreated and photobiotinylated DNA or RNA. After treatment with streptavidin and phenol, hybrids and single-stranded biotinylated material are sequestered to the aqueous/ organic interface while unhybridized material remains in the aqueous supematant. This procedure is simpler than published methods using either hydroxyapatite (HAP) chromatography (eg 1) or cupric-iminodiacetic acid agarose chromatography of biotinylated nucleic acid (2). The protocol given here is for subtraction of cDNA with RNA, but any combination of nucleic acid may be used.