Detection of nucleic acid reactions on bead arrays

• US 20050100893A1
• Filed: 10/15/2002
• Published: 05/12/2005
• Est. Priority Date: 04/20/1999
• Status: Abandoned Application

• Current Assignee: John Stuelpnagel, Kevin Gunderson, Mark S. Chee
• Original Assignee: John Stuelpnagel, Kevin Gunderson, Mark S. Chee
• Inventors: Gunderson, Kevin, Chee, Mark S., Stuelpnagel, John R.
• Application Number:
  US10/272,384
• Publication Number:
  US 20050100893A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2521/501 Ligase
• CPC Class Codes: C12Q 2565/518 characterised by the immobilisation of the nucleic acid sample or target
• CPC Class Codes: C12Q 2565/537 characterised by the capture oligonucleotide acting as a primer

Abstract

The present invention is directed to methods and compositions for the use of microsphere arrays to detect and quantify a number of nucleic acid reactions. The invention finds use in genotyping, i.e. the determination of the sequence of nucleic acids, particularly alterations such as nucleotide substitutions (mismatches) and single nucleotide polymorphisms (SNPs). Similarly, the invention finds use in the detection and quantification of a nucleic acid target using a variety of amplification techniques, including both signal amplification and target amplification. The methods and compositions of the invention can be used in nucleic acid sequencing reactions as well. All applications can include the use of adapter sequences to allow for universal arrays.

Claims

1-51. -51. (canceled)

52. A composition comprising a plurality of padlock probes for detecting a plurality of target sequences in a sample, wherein each target sequence comprises first and second target domains, and each of the padlock probes comprises:

a) a first probe sequence complementary to the first target domain;

b) a second probe sequence complementary to the second target domain;

c) a priming site that is identical for each of the plurality of probes; and

d) a restriction endonuclease site that is identical for each of the plurality of probes, wherein the restriction site occurs in a sequence intervening between the first probe sequence and the second probe sequence.

53. The composition of claim 52 wherein each padlock probe also comprises an adapter sequence.

54. The composition of claim 53 wherein the adapter sequence is unique to a specific combination of first and second probe sequences in a padlock probe.

55. The composition of claim 53, wherein the adapter sequence occurs in a sequence intervening between the first probe sequence and the second probe sequence.

56. The composition of claim 52, wherein the padlock probe has a first terminus comprising the first probe sequence and a second terminus comprising the second probe sequence.

57. The composition of claim 56, wherein the first terminus is hybridized to the first target domain and the second terminus is hybridized to the second target domain.

58. The composition of claim 57, wherein a circularized probe can be formed by ligation of the padlock probe.

59. The composition of claim 57, wherein a circularized probe can be formed by extension and ligation of the padlock probe.

60. The composition of claim 52, wherein the padlock probe is circularized.

61. The composition of claim 60, wherein the padlock probe has no terminus.

62. The composition of claim 52, wherein the first target domain comprises a single nucleotide polymorphism (SNP).