Patent ID: 6190865
Filing Date: 2001-02-20
Classification: C12Q

Abstract:
A method for characterizing a nucleic acid molecule comprising:a) synthesizing DNA in the presence of a reaction mixture comprising:(i) a nucleic acid template,(ii) a primer molecule that contains a nucleotide sequence at least part of which is sufficiently complementary to a portion of the template to hybridize therewith,(iii) an enzyme that extends the primer so that a DNA molecule may be synthesized,(iv) four canonical deoxynucleoside triphosphates, and(v) at least one non-canonical deoxynucleoside triphosphate, wherein the non-canonical deoxynucleoside triphosphate is one that is incorporated with fidelity into the synthesized DNA in place of only one canonical deoxynucleoside triphosphate;b) contacting the synthesized DNA with an N-glycosylase that excises a base portion of the non-canonical deoxynucleoside triphosphate from said synthesized DNA, whereby an abasic site is created;c) treating the DNA in such manner that said treatment results in breakage of a phosphodiester backbone at all abasic sites, whereby at least two DNA fragments are created; andd) separating the fragments according to size.