The present invention is directed to a method of sequencing a target
nucleic acid molecule having a plurality of bases. In its principle, the
temporal order of base additions during the polymerization reaction is
measured on a molecule of nucleic acid, i.e. the activity of a nucleic
acid polymerizing enzyme on the template nucleic acid molecule to be
sequenced is followed in real time. The sequence is deduced by
identifying which base is being incorporated into the growing
complementary strand of the target nucleic acid by the catalytic activity
of the nucleic acid polymerizing enzyme at each step in the sequence of
base additions. A polymerase on the target nucleic acid molecule complex
is provided in a position suitable to move along the target nucleic acid
molecule and extend the oligonucleotide primer at an active site. A
plurality of labelled types of nucleotide analogs are provided proximate
to the active site, with each distinguishable type of nucleotide analog
being complementary to a different nucleotide in the target nucleic acid
sequence. The growing nucleic acid strand is extended by using the
polymerase to add a nucleotide analog to the nucleic acid strand at the
active site, where the nucleotide analog being added is complementary to
the nucleotide of the target nucleic acid at the active site. The
nucleotide analog added to the oligonucleotide primer as a result of the
polymerizing step is identified. The steps of providing labelled
nucleotide analogs, polymerizing the growing nucleic acid strand, and
identifying the added nucleotide analog are repeated so that the nucleic
acid strand is further extended and the sequence of the target nucleic
acid is determined.