Patent ID: 8017319

Claim:
Method for sequencing nucleic acids comprising the steps: (a) providing an at least partially optically transparent support with a multitude of identical nucleic acid molecules immobilized thereon wherein the nucleic acid molecules carry several fluorescent marker groups, (b) progressive cleavage of individual nucleotide building blocks from the immobilized identical nucleic acid molecules, and (c) simultaneous determination of the base sequence of the plurality of nucleic acid molecules based on a time-dependent change in the fluorescence of the nucleic acid molecules and the time-dependent change in the fluorescence of the cleaved nucleotide building blocks caused by the cleavage of nucleotide building blocks, wherein the fluorescence is produced by beaming light into the support and generating an evanescent excitation field by internal reflection at the support surface in the area of the immobilized nucleic acid molecules, wherein the molecule sequencing is single molecule sequencing, wherein the cleavage of said multitude of individual nucleotide building blocks from a plurality of single immobilized nucleic acid molecules is measured and carried out in one single step by using a reagent selected from the group consisting of a single strand-specific exonuclease and a double strand-specific exonuclease, and wherein each nucleic acid molecule is measured separately and the measurements of each individual nucleic acid molecule are carried out in parallel by means of a detection matrix.