Composition and method for nucleic acid sequencing
First Claim
Patent Images
1. A polymerase-nucleic acid complex, said polymerase-nucleic acid complex comprising:
- a target nucleic acid, a family A or family B polymerase having a DNA binding cleft, and an attachment complex that attaches said polymerase on both sides of the DNA binding cleft and further wherein the attachment complex attaches the polymerase to a support,wherein said attachment complex irreversibly associates said target nucleic acid with said polymerase until replication is complete.
3 Assignments
0 Petitions
Accused Products
Abstract
The present invention provides compositions and methods for detecting incorporation of a labeled nucleotide triphosphate onto the growing end of a primer nucleic acid molecule. The method is used, for example, to genotype and sequence a nucleic acid. In a preferred embodiment, the method described herein detects individual NTP molecules.
-
Citations
52 Claims
-
1. A polymerase-nucleic acid complex, said polymerase-nucleic acid complex comprising:
-
a target nucleic acid, a family A or family B polymerase having a DNA binding cleft, and an attachment complex that attaches said polymerase on both sides of the DNA binding cleft and further wherein the attachment complex attaches the polymerase to a support, wherein said attachment complex irreversibly associates said target nucleic acid with said polymerase until replication is complete. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 45, 46, 47, 48, 49, 50, 51, 52)
-
-
27. A method for detecting incorporation of at least one NTP into a single primer nucleic acid molecule, said method comprising:
-
i. immobilizing onto a support a polymerase nucleic acid complex comprising a target nucleic acid, a primer nucleic acid which complements a region of the target nucleic acid, and at least one family A or family B polymerase having a DNA binding cleft, and an attachment complex that attaches said polymerase on both sides of the DNA binding cleft and further wherein the attachment complex attaches the polymerase to a support, wherein during replication, said attachment complex irreversibly associates said target nucleic acid with said polymerase until replication is complete; ii. contacting said immobilized complex with at least one type of labeled nucleotide triphosphate (NTP), wherein each NTP is labeled with a detectable label, and iii. detecting the incorporation of said at least one type of labeled NTP into a single molecule of said primer, while said at least one type of labeled NTP is in contact with said immobilized complex, by detecting the label of the NTP while said at least one type of labeled NTP is in contact with said polymerase nucleic acid complex. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44)
-
Specification