Cyclic single molecule sequencing process

A process for sequencing an individual nucleic acid, comprising the following steps:

• (a) providing a nucleic acid-synthesizing enzyme molecule, a circular nucleic acid template molecule, a primer annealed to said template, or capable of annealing to said template and fluorescence-labelled nucleotide building blocks,
• (b) generating a nucleic acid molecule complementary to the sequence of the circular nucleic acid template having incorporated said nucleic building blocks in a primer elongation catalyzed by a nucleic acid-synthesizing enzyme molecule,
• (c) contacting said generated nucleic acid molecule with a nucleic acid-degrading enzyme molecule and cleaving off individual nucleotide building blocks from said generated nucleic acid molecule in a nuclease digestion catalyzed by said nucleic acid-degrading enzyme molecule, and
• (d) determining the base sequence of said circular nucleic acid template molecule on the basis of the time-dependent fluorescence change, caused when nucleotide building blocks are incorporated during primer elongation and/or cleaved off during nuclease digestion,
• wherein the nucleic acid-synthesizing enzyme molecule is immobilized on a support by high-affinity interactions between partners of a specific binding pair, or adsorption, or covalent immobilization.