In Situ Hybridization Of Complementary Deoxyribonucleic Acid Probes To The 1448-1455 Region Of Escherichia coli 16s Ribosomal Subunits

Abstract

30S subunits were isolated from Escherichia coli ribosomes and purified 16S RNA was subsequently extracted. An eight-base DNA oligomer [d(CCCGAAGG)] was hybridized in situ to the 1448-1455 region of the 16S rRNA. The cDNA-rRNA hybrid was digested with the RNA specific enzyme RNase H. The clipped RNA fragments were then separated by polyacrylamide gel electrophoresis. The desired RNA band was visualized by UV shadowing, eluted from the gel and sequenced to verify the specificity of the probe binding. The results suggest that the cDNA and rRNA were binding, but proof that it was specific to the 1448-1455 region is still lacking.