Methods and compositions for incorporating nucleotides

摘要

The invention provides methods and compositions, including, without limitation, algorithms, computer readable media, computer programs, apparatus, and systems for determining the identity of nucleic acids in nucleotide sequences using, for example, data obtained from sequencing by synthesis methods. The methods of the invention include correcting one or more phenomena that are encountered during nucleotide sequencing, such as using sequencing by synthesis methods. These phenomena include, without limitation, sequence lead, sequence lag, spectral crosstalk, and noise resulting from variations in illumination and/or filter responses.

主权项

1. A method for determining an identity of a nucleotide at a position in a nucleotide sequence, comprising: a) providing primers, nucleic acid template molecules having a nucleotide sequence, polymerase, and a mixture of labeled and non-labeled reversible terminators, said labeled reversible terminators comprising a label linked to a nucleotide analogue, said label providing a signal, said signal having an intensity; b) incorporating, with a polymerase, a labeled reversible terminator from said mixture into a primer bound to a nucleic acid template molecule at a position corresponding to a complementary base in said template molecule; c) measuring the intensity of the signal of said label of said incorporated labeled reversible terminator; and d) determining the identity of the nucleotide at said position.