Efficient base determination in sequencing reactions
First Claim
1. A method of determining the identification of a first nucleotide at a detection position of a target sequence comprising a plurality of detection positions, said method comprising:
- (a) providing a surface comprising a plurality of concatemers, wherein each concatemer comprises a plurality of monomers and each monomer comprises;
i) a first target domain of said target sequence comprising a first set of target detection positions;
ii) at least a first adaptor comprising;
1) a first anchor site; and
2) a second adjacent anchor site;
b) hybridizing a first anchor probe to said first anchor site;
c) hybridizing a second anchor probe to said second anchor site, wherein said second anchor probe also hybridizes to sequences outside said second anchor site;
d) hybridizing at least a first sequencing probe to said first target domain, wherein said first sequencing probe comprises;
i) a first probe domain complementary to said target domain;
ii) a unique nucleotide at a first interrogation position; and
iii) a label;
under conditions wherein if said unique nucleotide is complementary to said first nucleotide, said sequencing probe hybridizes to said concatemer; and
e) ligating said anchor probes and said sequencing probe;
f) identifying said first nucleotide.
1 Assignment
0 Petitions
Accused Products
Abstract
The present invention is directed to compositions and methods for nucleic acid identification and detection. Compositions and methods of the present invention include extracting and fragmenting target nucleic acids from a sample, using the fragmented target nucleic acids to produce target nucleic acid templates and subjecting those target nucleic acid templates to amplification methods to form nucleic acid nanoballs. The invention also includes methods of detecting and identifying sequences using various sequencing applications, including sequencing by ligation methods.
-
Citations
14 Claims
-
1. A method of determining the identification of a first nucleotide at a detection position of a target sequence comprising a plurality of detection positions, said method comprising:
-
(a) providing a surface comprising a plurality of concatemers, wherein each concatemer comprises a plurality of monomers and each monomer comprises; i) a first target domain of said target sequence comprising a first set of target detection positions; ii) at least a first adaptor comprising; 1) a first anchor site; and 2) a second adjacent anchor site; b) hybridizing a first anchor probe to said first anchor site; c) hybridizing a second anchor probe to said second anchor site, wherein said second anchor probe also hybridizes to sequences outside said second anchor site; d) hybridizing at least a first sequencing probe to said first target domain, wherein said first sequencing probe comprises; i) a first probe domain complementary to said target domain; ii) a unique nucleotide at a first interrogation position; and iii) a label; under conditions wherein if said unique nucleotide is complementary to said first nucleotide, said sequencing probe hybridizes to said concatemer; and e) ligating said anchor probes and said sequencing probe; f) identifying said first nucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
-
Specification