Polymorphism detection

• US 6,586,186 B2
• Filed: 08/24/2001
• Issued: 07/01/2003
• Est. Priority Date: 11/29/1995
• Status: Expired due to Fees

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Sapolsky, Ronald, Lipshutz, Robert J., Ghandour, Ghassan
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US09/939,119
• Publication Number:
  US 20020098496A1
• Time in Patent Office: 676 Days
• Field of Search: 435/6, 536/22.1, 536/24.2, 536/24.3, 536/24.32
• US Class Current: 435/6.11
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00695 Synthesis control routines, e.g. using computer programs
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

The present invention generally provides a rapid efficient method for analyzing polymorphic or biallelic markers, and arrays for carrying out these analyses. In general, the methods of the present invention employ arrays of oligonucleotide probes that are complementary to target nucleic acids which correspond to the marker sequences of an individual. The probes are typically arranged in detection blocks, each block being capable of discriminating the three genotypes for a given marker, e.g., the heterozygote or either of the two homozygotes. The method allows for rapid, automatable analysis of genetic linkage to even complex polygenic traits.

Claims

1. An array of oligonucleotide probes for detecting a polymorphism in a target nucleic acid sequence using Principal Component Analysis, said array comprising at least one detection block of probes, said detection block including a first group of probes that are complementary to said target nucleic acid sequence except that the group of probes includes all possible monosubstitutions of positions in said sequence that are within n bases of a base in said sequence that is complementary to said polymorphism, wherein n is from 0 to 5, and a second and third group of probes complementary to marker-specific regions upstream and downstream of the polymorphism in the target nucleic acid sequence, wherein the third group of probes differs from the second set of probes at single bases corresponding to known mismatch positions.

2. The array of claim 1, wherein the polymorphism is identified as a result of Principal Component Analysis of hybridization intensities of the array of probes.

3. The array of claim 1, wherein at least two alleles of the polymorphism are known.

4. The array of claim 1, wherein said first group of probes comprises a plurality of different probes that are complementary to overlapping portions of said target nucleic acid sequence.

5. The array of claim 1, wherein the monosubstitutions occur at a plurality of distances from a 3′

end of said probes.

6. The array of claim 1, wherein said detection block includes between about 8 and 88 different probes.

7. The array of claim 1, comprising between 1 and 1,000 different detection blocks, each of said detection blocks including probes complementary to different polymorphisms in said target nucleic acid sequence.

8. An array of oligonucleotide probes for detecting a polymorphism in a target nucleic acid sequence, said array comprising:

a first group of probes that are complementary to said target nucleic acid sequence except that the group of probes includes all possible monosubstitutions of positions in said sequence that are within n bases of a base in said sequence that is complementary to said polymorphism; and

a second and third group of probes complementary to marker-specific regions upstream and downstream of the polymorphism in the target nucleic acid sequence, wherein the third group of probes differs from the second set of probes at single bases corresponding to known mismatch positions.

9. The array of claim 8, wherein at least two alleles of the polymorphism are known.

10. The array of claim 8, wherein said first groups of probes comprises a plurality of different probes that are complementary to overlapping portions of said target nucleic acid sequence.

11. The array of claim 8, wherein the monosubstitutions occur at a plurality of distances from a 3′

end of said probes.

12. The array of claim 8, wherein said detection block includes between about 8 and 88 different probes.

13. The array of claim 8, comprising between 1 and 1,000 different detection blocks, each of said detection blocks including probes complementary to different polymorphisms in said target nucleic acid sequence.