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.