SHORT CYCLE METHODS FOR SEQUENCING POLYNUCLEOTIDES
The invention provides methods for sequencing a polynucleotide comprising stopping an extension cycle in a sequence by synthesis reaction before the reaction has run to near or full completion.
1 - 26 . (canceled)
27 . A method for sequencing a nucleic acid template, the method comprising the steps of:
(a) extending a primer hybridized to a nucleic acid template in a primer/template duplex by exposing said primer to (i) a polymerase capable of catalyzing nucleotide addition to said primer, and (ii) one or more extendible nucleotides each comprising a label, under conditions wherein said polymerase extends said primer by incorporation of a nucleotide;
(b) identifying said nucleotide incorporated into said primer;
(c) repeating said extending and said identifying steps to determine the nucleotide sequence of the nucleic acid molecule.
28 . The method of claim 27 , wherein said one or more extendible nucleotides comprises a single type of nucleotide.
29 . The method of claim 27 , wherein said detectable label comprises an optically detectable label.
30 . The method of claim 29 , wherein said optically detectable label comprises a fluorescent label.
31 . The method of claim 29 , wherein said optically detectable label comprises a chemiluminescent label.
32 . The method of claim 27 , wherein said identifying comprising detecting label on said nucleotide incorporated into said primer.
33 . The method of claim 27 , further comprising a step wherein unbound nucleotides are removed after said extending and prior to said identifying.