[A new method of covalent immobilization of oligodeoxyribonucleotides on nylon membranes for hybridization with...

A new method of covalent immobilization of oligodeoxyribonucleotides on nylon membranes which contain surface amino groups was developed. The method consists in condensation between the amino group of the membrane and the carboxyl group of modified oligonucleotide by means of 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide. The carboxyl group was introduced into the oligonucleotide by means of postsynthetic attachment of peptide (reduced glutathione) at the terminal phosphate group of the oligonucleotide, using the N-hydroxybenzotriazole method of phosphate activation. Membranes containing a covalently immobilized 23-membered oligonucleotide were tested in hybridization with complementary oligonucleotide, and with single-stranded DNA of bacteriophage M13 which has a complementary sequence. The method of covalent immobilization is very simple and convenient. The membranes with covalently immobilized oligonucleotides may be used not only in hybridization analysis, but also for purification of nucleic acids and proteins which recognize nucleotide sequences and in sense biotechnology.