| abstract |
The invention relates an oligonucleotide probe reagent consisting of a nylon membrane with a high density of anionic carboxyl groups to which membrane at least one oligonucleotide probe containing a 5' amine is covalently bound via an amide bond and to a method for attachment of the oligonucleotide probe to the nylon membrane. The single, 5' end attachment of the oligonucleotides to the membrane surface leaves the probe free to interact with complementary sequences, thus increasing the hybridization efficiency relative to probes attached by methods in which heat or ultraviolet light is used for immobilization. The simplicity and reproducibility of this method and the sensitivity attained when using the reagents produced by the method are ideal for application of the method and reagents to the diagnosis of infectious and genetic diseases, the analysis of mutations in neoplasias, performance of HLA typing, and other areas. |