METHOD FOR THE AMPLIFICATION AND OPTIONAL CHARACTERISATION OF NUCLEIC ACIDS

摘要

A method for the amplification of a template nucleic acid comprises simultaneously carrying out the steps of reacting a nucleic acid primer with said template nucleic acid, normal DNA precursor nucleotides, at least one modified DNA precursor nucleotide and a DNA polymerase so as to obtain an extended nucleic acid primer, said nucleic acid primer remaining bound to said template; cleaving the modified base-containing extended nucleic acid primer so as to generate a free 3'-OH terminus that is extensible by said DNA polymerase; and repeating steps i) and ii) on DNA fragments thereby generated. The modified precursor nucleotide may be a substrate for a DNA glycosylase or recognised by a 3'-endonulcease and determines the cleavage of the DNA and the site of the cleavage accordingly. The method has significant advantages over existing technologies in that it is more versatile and more flexible with respect to providing a single high throughput process that can be easily adapted to multiple different formats in the fields of DNA detection, quantitation and characterisation.