Complex oligonucleotide primer mix

• US 20090036664A1
• Filed: 07/31/2007
• Published: 02/05/2009
• Est. Priority Date: 07/31/2007
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Peter, Brian Jon
• Application Number:
  US11/888,388
• Publication Number:
  US 20090036664A1
• Time in Patent Office: Days
• US Class Current: 536/25.300
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical

Abstract

The invention generally relates to a complex mixture of oligonucleotide primers and/or probes. Another aspect of the invention includes a method of selective priming of a target nucleic acid.

Claims

1. A composition comprising a plurality of isolated oligonucleotides at least some of which are nonidentical in sequence, length or both, wherein each of the oligonucleotides comprises:

a) a defined sequence of at least 16 nucleotides, wherein at least a portion of the sequence is complementary to a target nucleic acid,

b) at least one cleavage site, and optionally

c) at least two different subsequences separated by at least one of the cleavage site sequences, wherein each of the subsequences of each oligonucleotide binds to a different site in the target nucleic acid or each of the subsequences binds to a different target nucleic acid.

2. The composition of claim 1, wherein the target nucleic acid is genomic DNA or a chromosomal fragment.

3. The composition of claim 1, wherein each oligonucleotide or subsequence thereof has a sequence that does not bind to a repeat sequence in the target nucleic acid.

4. The composition of claim 1, wherein each oligonucleotide or subsequence thereof does not bind to a site of the target nucleic acid of at least 200 bp comprising at least 50% GC content and an observed/expected CpG ratio greater than 0.6.

5. The composition of claim 1, wherein each oligonucleotide comprises at least three different subsequences, each of the subsequences separated by at least a cleavage site, wherein each of the subsequences binds to a different site on the target nucleic acid.

6. The composition of claim 1, wherein the cleavage site is specifically cleaved by light, a chemical, or a restriction enzyme.

7. The composition of claim 1, wherein each oligonucleotide comprises at least 100 nucleotides.

8. The composition of claim 1, wherein the oligonucleotides are labelled.

9. A method for preparing a plurality of primers, comprising:

a) selecting at least one target nucleic acid;

b) identifying a sequence for each of a plurality of oligonucleotides, wherein each sequence comprises

i) a defined sequence of at least 100 nucleotides wherein at least a portion of the sequence is complementary to a target nucleic acid, and

ii) at least one cleavage site;

c) synthesizing each of the oligonucleotides at a different address on the substrate; and

d) cleaving each of the oligonucleotides from the array.

10. The method of claim 9, wherein each oligonucleotide is cleaved from the substrate and cleaved into at least two subsequences in step (d).

11. The method of claim 9, wherein the target nucleic acid is genomic DNA.

12. The method of claim 9, wherein each oligonucleotide has a sequence that does not bind to a repeat sequence in the target nucleic acid.

13. The method of claim 9, wherein each oligonucleotide does not bind to a site of the target nucleic acid of at least 200 bp comprising at least 50% GC content and an observed/expected CpG ratio greater than 0.6.

14. The method of claim 9, wherein a cleavage site is a phosphoramidite spacer.

15. The method of claim 9, wherein each oligonucleotide comprises at least three different subsequences, each of the subsequences separated by at least a cleavage site, wherein each of the subsequences binds to a different site on the target nucleic acid.

16. The method of claim 9, wherein the oligonucleotide comprises at least two cleavage sites.

17. The method of claim 9, wherein each oligonucleotide comprises at least 200 nucleotides.

18. The method of claim 9, wherein at least 50% of the oligonucleotides or subsequences thereof bind to a different site in the same target nucleic acid, wherein the target nucleic acid is at least 500 base pairs in length.

19. The method of claim 9 further comprising labeling the oligonucleotides.

20. The method of claim 19, wherein the labeling comprises fluorescent labels.