Hydroxymethyl Linkers For Labeling Nucleotides
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.
1 . A method of incorporating labeled nucleotides into nucleic acid, comprising:
a) providing a plurality of nucleic acid template molecules, a polymerase, and a plurality of nucleotide analogues wherein each nucleotide analogue is labeled with a unique label and contains a removable chemical moiety capping the 3′-OH group, wherein the label is attached to the base through a hydroxymethyl group;
b) incorporating a first nucleotide analogue with said polymerase; and
c) detecting the label of the incorporated nucleotide analogue.