Methods for sequence determination
0 Assignments
0 Petitions
Accused Products
Abstract
A sequencing methodology is disclosed that allows a single DNA or RNA molecule or portion thereof to be sequenced directly and in substantially real time. The methodology involves engineering a polymerase and/or dNTPs with atomic and/or molecular tags that have a detectable property that is monitored by a detection system.
148 Citations
51 Claims
-
1. (canceled)
-
2. (canceled)
-
3. (canceled)
-
4. (canceled)
-
5. (canceled)
-
6. (canceled)
-
7. (canceled)
-
8. (canceled)
-
9. (canceled)
-
10. (canceled)
-
11. (canceled)
-
12. (canceled)
-
13. (canceled)
-
14. (canceled)
-
15. (canceled)
-
16. (canceled)
-
17. (canceled)
-
18. (canceled)
-
19. (canceled)
-
20. (canceled)
-
21. (canceled)
-
22. (canceled)
-
23. (canceled)
-
24. (canceled)
-
25. (canceled)
-
26. (canceled)
-
27. (canceled)
-
28. (canceled)
-
29. (canceled)
-
30. (canceled)
-
31. (canceled)
-
32. A method of sequencing an individual nucleic acid molecule or numerous individual molecules in parallel comprising the steps of:
-
immobilizing a member of the replication complex comprising a polymerizing agent including a polymerizing agent tag a primer and a nucleic acid template sufficiently spaced apart to allow resolvable detection of one or more complexes on a solid support;
contacting the immobilized member with the other members to form bound replication complexes;
incubating the bound replication complexes with monomer for the polymerizing agent, where at least one monomer type includes monomer tag covalently bonded to a portion of the monomer that is released when the tagged monomer is incorporated;
detecting with a detector a change in a detectable property of at least one of the tags during one monomer incorporation event or a plurality of monomer incorporation events; and
relating the changes in the detectable property to an identity of the incorporated monomer or identities of the incorporated plurality of monomer and correlating the identities to one or a sequence of nucleoside in an unknown nucleic acid. - View Dependent Claims (35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51)
-
-
33. (canceled)
-
34. (canceled)
Specification