Patent ID: 8715938

Claim:
A method for determining a sequence of a template polynucleotide, the method comprising: a) hybridizing a first primer to the template polynucleotide; b) contacting at least one labeled reversible di-nucleotide terminator and a DNA polymerase with the first primer to extend the first primer along the template by polymerase-mediated polymerization of the labeled reversible di-nucleotide terminator to form a first extension product, wherein the labeled reversible di-nucleotide terminator includes a first nucleotide connected to a second nucleotide and includes a detectable moiety linked to the first nucleotide and includes a blocking moiety linked to the second nucleotide; c) detecting the detectable moiety of the polymerized labeled reversible di-nucleotide terminator of step (b); d) identifying a probe family to which the polymerized labeled reversible di-nucleotide terminator in step (b) belongs, the probe family having a plurality of labeled reversible di-nucleotide terminators of varying sequence each having the same type of a detectable moiety; e) removing the blocking group on the polymerized labeled reversible di-nucleotide terminator of step (b) to generate a free 3′ OH group; f) repeating steps (b), (c), (d), and optionally (e), to generate a first accumulated extension product; g) generating a set of candidate ordered series of the probe families which correlate with an ordered series of the polymerized labeled reversible di-nucleotide terminators in steps (b), (c) and (d); and h) determining the sequence of the template polynucleotide based at least in part on the first set of candidate ordered series of the probe families.