Methods and compositions for inhibiting undesired cleaving of labels
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 composition comprising a cleaving agent scavenger and one or more nucleotide analogues in solution, said scavenger comprising cystamine, said analogues labeled with a unique label attached through a cleavable linker.
2. The composition of claim 1 , wherein said unique label is attached through a cleavable disulfide linker.
3. The composition of claim 1 , wherein said unique label is attached through a cleavable azido linker.
4. The composition of claim 1 , wherein said one or more nucleotide analogues comprise a removable chemical moiety capping the 3′-OH.
5. The composition of claim 4 , wherein said chemical moiety comprises a disulfide bond.
6. The composition of claim 4 , wherein said chemical moiety comprises an azido group.
7. The composition of claim 4 , wherein said chemical moiety comprises an azidomethyl ether.
8. The composition of claim 1 , further comprising a polymerase.
9. A system comprising reservoirs in fluid communication with a reaction chamber, at least one of said reservoirs comprising a solution comprising a cleaving agent scavenger and one or more nucleotide analogues, said scavenger comprising cystamine, said analogues labeled with a unique label attached through a cleavable linker.
10. The system of claim 9 , wherein said unique label is attached through a cleavable disulfide linker.
11. The system of claim 9 , wherein said unique label is attached through a cleavable azido linker.
12. The system of claim 9 wherein said one or more nucleotide analogues comprise a removable chemical moiety capping the 3′-OH.
13. The system of claim 12 , wherein said chemical moiety comprises a disulfide bond.
14. The system of claim 12 , wherein said chemical moiety comprises an azido group.
15. The system of claim 12 , wherein said chemical moiety comprises an azidomethyl ether.