NONRADIOACTIVE METHODS OF LABELING AND DETECTION OF NUCLEIC-ACIDS DURING HYBRIDIZATION

Hybridization of nucleic acids represents one of the important methods of molecular biology for proving the presence of some sequences of nucleotides in the DNA and RNA molecules by means of radioactive and non-radioactive labelled probes. The review article stresses possible methods of obtaining non-radioactive probes by means of binding biotin derivatives, such as photobiotin, biocytin, biotinhydrazide, biotin-11-dUTP and other onto single stranded or double stranded DNA, RNA and also synthetic oligonucleotides. Mention is also made of the possibilities of the detection of hybridization of nucleic acids by probes labelled with biotin derivatives, both enzymatic and chemiluminescence and also detection with antibiotin antibodies. Among other methods of labelling and detection of nucleic acids during hybridization, acetylaminofluoren, dioxigenin, piezoelectric resonance and direct binding of the enzyme onto DNA are mentioned. In conclusion the sensitivities of individual types of labelling are compared in relation to the radioactively labelled hybridization with P-32 and S-35.