| abstract |
Method for mass spectrometric determination of known mutations in genomic DNA, consisting of the following steps: (1) an amount of DNA is provided which contains the mutation, (2) primers and a set of modified, non-terminating nucleoside triphosphates (NTP) and terminating nucleoside triphosphates for primer extension are added, the modifications of the nucleoside triphosphates allowing the neutralization of the primer extension products in step (5), and " Charge tags "are attached either to the primers or to the terminating nucleoside triphosphates, (3) the primers are hybridized to a suitable DNA strand and extended in an enzymatic copying reaction, (4) at least a defined part of each of the primers is split off or removed from the extended primer product chains, (5) the remaining product chains, with the exception of the “charge tags”, are chemically neutralized, the modified sites of the nucleobases being used for neutralization and (6) the mass of the product chains is determined by mass spectrometry and the proven masses are assigned to the wild type or known mutants, without first ... |