Directed enrichment of genomic DNA for high-throughput sequencing
First Claim
1. A microarray comprising a plurality of oligonucleotide primer pairs, wherein each oligonucleotide primer pair comprises a first oligonucleotide primer and a second oligonucleotide primer, each primer comprising at least one cleavable linkage for releasing the plurality of oligonucleotide primer pairs from the microarray.
3 Assignments
0 Petitions
Accused Products
Abstract
The present invention provides microarrays of oligonucleotide primer pairs, and in particular, microarrays of primers that comprise at least one cleavable linkage. Also provided are methods to capture oligonucleotide primer pairs from one or more microarrays, and methods to use the captured oligonucleotide primer pairs, such as for amplification of a target polynucleotide sequence. In addition, methods of using a microarray to isolate, purify and/or amplify a target polynucleotide are provided.
-
Citations
42 Claims
- 1. A microarray comprising a plurality of oligonucleotide primer pairs, wherein each oligonucleotide primer pair comprises a first oligonucleotide primer and a second oligonucleotide primer, each primer comprising at least one cleavable linkage for releasing the plurality of oligonucleotide primer pairs from the microarray.
-
14. A method for producing a microarray, wherein a plurality of oligonucleotide primer pairs are synthesized on the microarray, each oligonucleotide primer pair is present at a discrete location on the microarray and comprises a first primer and a second primer, and the first primer and second primer each comprise at least one cleavable linkage, comprising:
-
a) providing a microarray comprising a plurality of discrete locations, wherein each discrete location comprises a first primer synthesis site and a second primer synthesis site;
b) capping each second primer synthesis site with a blocking group;
c) synthesizing a first primer comprising at least one cleavable linkage at each first primer synthesis site on the microarray, thereby producing a plurality of first primers on the microarray;
d) capping the plurality of first primers, thereby preventing further synthesis of each first primer;
e) uncapping each second primer synthesis site; and
f) synthesizing a second primer comprising at least one cleavable linkage at each second primer synthesis site on the microarray, thereby producing a plurality of second primers on the microarray;
thereby producing a microarray, wherein a plurality of oligonucleotide primer pairs are synthesized on the microarray. - View Dependent Claims (15)
-
-
16. A method for capturing a plurality of oligonucleotide primer pairs on a capturing means in a capturing support, wherein each oligonucleotide primer pair is present at a discrete location on the capturing support, comprising:
-
a) embedding each oligonucleotide primer pair present at a discrete location on a microarray into a capturing support, wherein each oligonucleotide primer comprises at least one first cleavable linkage and the capturing support comprises a capturing means for capturing the oligonucleotide primer pairs;
b) capturing the oligonucleotide primer pairs on the microarray by the capturing means; and
c) separating the oligonucleotide primer pairs from the microarray by cleaving the at least one first cleavable linkage;
thereby capturing a plurality of oligonucleotide primer pairs on a capturing means in a capturing support, wherein each oligonucleotide primer pair is present at a discrete location in the capturing support on a capturing means. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33)
-
-
17-25. -25. (canceled)
-
34-35. -35. (canceled)
-
36. A method for capturing a plurality of oligonucleotide primer pairs on a capturing support, wherein each oligonucleotide primer pair comprises a first primer and a second primer, and wherein each first primer is captured from a first microarray and each second primer is captured from a second microarray, comprising:
-
a) embedding a first microarray comprising a plurality of first primers, wherein each first primer is located on the first microarray at a discrete location, into a capturing support under conditions in which each of the first primers are captured in the capturing support and the discrete location of each of the first primers is maintained in the capturing support, and wherein each first primer comprises at least one first cleavable linkage located at the microarray surface;
b) separating the first primers from the first microarray by cleaving the at least one first cleavable linkage in the first primers;
c) removing the first microarray from the capturing support;
d) embedding a second microarray comprising a plurality of second primers, wherein each second primer is located on the second microarray at a discrete location, into the capturing support under conditions in which each of the second primers are captured in the capturing support and the discrete location of each of the second primers is maintained in the capturing support, wherein the discrete location of each of the second primers overlaps with the discrete location of each of the first primers, and wherein each second primer comprises at least one second cleavable linkage;
e) separating the second primers from the second microarray by cleaving the at least one second cleavable linkage in each of the second primers;
thereby capturing a plurality of oligonucleotide primer pairs on a capturing support, wherein each oligonucleotide primer pair comprises a first primer and a second primer, and wherein each first primer is captured from a first microarray and each second primer is captured from a second microarray. - View Dependent Claims (37, 38, 39, 40, 41)
-
-
42-97. -97. (canceled)
Specification