Methods for production of arrays with modified oligonucleotide and polynucleotide compositions

• US 6,844,151 B1
• Filed: 09/29/1999
• Issued: 01/18/2005
• Est. Priority Date: 09/29/1999
• Status: Expired due to Fees

• Current Assignee: Lakewood Amedex Inc.
• Original Assignee: Oligos Etc., Inc.
• Inventors: Dale, Roderic M. K.
• Primary Examiner(s): Allen, Marianne P.
• Application Number:
  US09/408,761
• Time in Patent Office: 1,938 Days
• Field of Search: 435/6, 435/5, 536/24.5, 536/23.1, 536/24.3, 536/25.4
• US Class Current: 435/6.14
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence

Abstract

The present invention provides methods for producing arrays having associated modified nucleic acid structures, e.g. acid stable and/or end-blocked nucleic acids such as 2′-O—R oligonucleotides. In one embodiment, arrays produced using the methods of the invention exhibit an increased binding affinity with complementary nucleic acids, and in particular with complementary RNA. In another embodiment, the associated nucleic acids of the array produced using the methods of the invention exhibit substantial acid resistance, allowing the arrays to be treated with low pH solutions. In another embodiment, the modified associated nucleic acids of the array produced using the methods of the invention exhibit substantial resistance to nuclease degradation.

Claims

1. An array comprising:

(A) a support surface; and

(B) a plurality of modified oligonucleotide compositions, each composition comprising a plurality of oligonucleotides stably associated with a distinct area of the support surface, wherein the oligonucleotides of each composition are characterized by;

(1) independently having a length of from about 20 to about 300 nucleotides;

(2) having internucleotide linkages selected from the group consisting of 2′

-O-alkyl, 2′

-O-alkyl-n(O-alkyl), combinations thereof;

(3) having a binding affinity to a complementary sequence greater than the corresponding binding affinity of a non-modified oligonucleotide having the same sequence; and

(4) having a pH stability of at least one hour at 37°

C. at a pH in a range of about 0.5 to 6.

2. The array of claim 1, wherein the modified oligonucleotides further comprise an end block at the 3′

end and exhibit exonuclease resistance of at least twice that of a naturally occurring oligonucleotide having the same sequence and number of bases.

3. The array of claim 1, wherein the modified oligonucleotides further comprise an end block at the 5′

end and exhibit exonuclease resistance of at least twice that of a naturally occurring oligonucleotide having the same sequence and number of bases.

4. An array of claim 1, wherein the modified oligonucleotides of each distinct area of the array exhibit substantially the same T_m.

5. The array of claim 1, wherein the modified oligonucleotides of a distinct area are selectively designed to hybridize to RNA.

6. The array of claim 1, wherein the modified oligonucleotides of a distinct area are selectively designed to hybridize to DNA.

7. The array of claim 1, wherein the modified oligonucleotides are further characterized by modification of at least 25% of their internucleoside linkages.

8. The array of claim 1, wherein said modified oligonucleotides have an average length of from about 100 to about 200 nucleotides.

9. The array of claim 1, wherein oligonucleotide sequences of the oligonucleotides of an oligonucleotide composition on the array differs from those of the oligonucleotides of any other oligonucleotide composition on said array.

10. The array of claim 1, wherein each oligonucleotide composition comprises a population of identical oligonucleotides.

11. The array of claim 1, wherein each oligonucleotide composition comprises a plurality of oligonucleotides that bind to a particular nucleic acid.

12. The array of claim 1, wherein the number of oligonucleotide compositions on said ranges from about 2 to about 10⁹ oligonucleotides.