Method for sequencing polynucleotides

摘要

A method for obtaining a candidate nucleotide sequence S indicative of a sequence of a target polynucleotide molecule that produces a hybridization signal I({right arrow over (x) upon incubation with a polynucleotide {right arrow over (x for each polynucleotide {right arrow over (x in a set E of polynucleotides. For each polynucleotide {right arrow over (x in the set E of polynucleotides, a probability P₀({right arrow over (x) of the hybridization signal I({right arrow over (x) when the sequence {right arrow over (x is not complementary to a subsequence of T and a probability P₁({right arrow over (x) of the hybridization signal when the sequence {right arrow over (x is complementary to a subsequence of T are obtained; so as to obtain a probabilistic spectrum (PS) of T. A score is then assigned to each of a plurality of candidate nucleotide sequences that is being based upon the probabilistic spectrum and upon a reference nucleotide sequence H. A candidate nucleotide sequence having an essentially maximal score is selected and one or more low confidence intervals and one or more reliable intervals in the selected candidate nucleotide sequence are identified. For each low confidence interval detected in the selected candidate nucleotide sequence, a score is assigned to each of a plurality of candidate nucleotide sequences of the low confidence region, where the score is based upon a probabilistic spectrum obtained by filtering from the PS signals the signals present in the reliable regions; and upon an interval of the reference nucleotide sequence H homologous with the low confidence interval. A candidate nucleotide sequence having an essentially maximal score is then selected. A revised candidate sequence S' is then obtained indicative of the sequence of the target polynucleotide molecule T by substituting the sequence of the low confidence region in the candidate sequence S with the selected candidate sequence.