A METHOD OF OLIGONUCLEOTIDE IMMOBILIZATIO N ON CARBOXYL-CONTAINING NYLON MEMBRANES FOR HYBRIDIZATION WITH NUCLEIC-ACIDS

A simple and efficient method of the covalent immobilization of oligon ucleotides on carboxyl-containing nylon membranes was proposed. The me thod is based on the reaction between a postsynthetically introduced a minoalkyl group of an oligonucleotide and membrane carboxyl catalyzed by a water soluble N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide. Jud ging from the mechanism of carboxyl group activation by the carbodiimi de, optimal conditions of the immobilization were selected, which made it possible to increase the immobilization efficiency and carrier cap acity as compared with those described previously. Using a 23-unit mem brane-immobilized oligonucleotide as an example, the oligonucleotides immobilized were shown to be capable of hybridizing with complementary sequences.