ABSTRACT
This chapter outlines an experimental approach to study the influence of bioregulators on gene expression. It discusses pertinent techniques involving isolation of total ribonucleic acid (RNA), fractionation of RNA, in vitro translation, one-and two-dimensional gel electrophoresis, immunoprecipitation, complementary deoxyribonucleic acid (DNA) synthesis, plasmid probe construction and screening, and northern blotting. The methodology involves binding one clone DNA to diazobenzyloxymethyl paper and screening the others via molecular hybridization. The probe DNA can be used to identify the complementary sequence in the partial restriction digests of cellular DNA, and the results interpreted in terms of number of genes in the genome. The analytical procedures allow one to characterize the protein and ribonucleic acid profiles of a tissue using methods of molecular biology including immunology and recombinant DNA techniques. Cetyltrimethylammonium bromide has been shown to preferentially precipitate RNA from DNA in mixtures of partially purified total cellular nucleic acid.
