Method for detecting DNA base sequence modifications in a sample DNA comprises amplifying the sample DNA by PCR, splitting the PCR product by uracil-specific nuclease and analyzing the fragments by mass-spectrometer

摘要

Method for detecting DNA base sequence modifications in a sample DNA comprises: amplifying the sample DNA in a PCR; splitting the PCR product into fragments by a uracil-specific nuclease; analyzing the fragments through mass-spectrometer; detecting the presence of DNA base sequence modifications by comparing the mass-spectrometrically determined fragment masses with the fragment masses determined for the presence of such DNA base sequence modifications. Method for detecting DNA base sequence modifications in a sample DNA comprises: amplifying the sample DNA in a PCR, where at least one of the employed primers contains at least at a position, a uracil instead of a base, preferably instead of a thymine, the primers used for the PCR bind to the segments of the sample DNA flanking the DNA base sequence modification to be detected, the PCR is accomplished in the presence of deoxyuridine triphosphate, deoxyadenosine triphosphate, deoxyguanosine triphosphate and deoxycytidine triphosphate; and a PCR product with a length of 33-60 base pairs is formed; splitting the PCR product into fragments by a uracil-specific nuclease; analyzing the fragment through mass-spectrometer; detecting the presence of a DNA base sequence modification by comparing the mass-spectrometrically determined fragment masses with the fragment masses determined for the presence of such DNA base sequence modifications.