Process for synthesizing oligonucleotides with random codons

• US 5,264,563 A
• Filed: 12/14/1992
• Issued: 11/23/1993
• Est. Priority Date: 08/24/1990
• Status: Expired due to Term

• Current Assignee: Ixsys, Inc. (Integral Partners LLC)
• Original Assignee: Ixsys, Inc. (Integral Partners LLC)
• Inventors: Huse, William D.
• Primary Examiner(s): Rollins, John W.
• Assistant Examiner(s): Kunz, Gary L.
• Application Number:
  US07/990,355
• Time in Patent Office: 344 Days
• Field of Search: 536/25.3, 536/25.31, 536/25.33, 536/25.34
• US Class Current: 536/25.3
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

The invention provides a method of synthesizing oligonucleotides having random tuplets using individual monomers. The steps consist of: (1) sequentially coupling monomers on separate supports to form at least two different tuplets, the coupling is performed in separate reaction vessels; (2) mixing the supports from the reaction vessels; (3) dividing the mixed supports into two or more separate reaction vessels; and (4) repeating steps (1) through (3) one or more times in the reaction vessels of step (3), wherein the last step ends at step (2). Additionally, the oligonucleotides can be cleaved from the supports.

Claims

1. A process of synthesizing oligonucleotides having random triplet codons using individual monomers, said process comprising:

(1) sequentially coupling monomers on separate supports to form at least two different triplet codons, said coupling being performed in separate reaction vessels;

(2) mixing the supports from the separate reaction vessels;

(3) dividing the mixed supports into two or more separate reaction vessels; and

(4) repeating steps (1) through (3) one or more times in the reaction vessels of step (3), wherein the last step ends at step (2).

2. The process of claim 1 further comprising (5) cleaving said oligonucleotides with random triplet codons from said support.

3. The process of claim 1, wherein the number of triplet codons to be randomized is between two and twenty.

4. The process of claim 1, wherein the number of reaction vessels is equal to the number of triplet codons randomized.

5. The process of claim 4, wherein each triplet codon randomized is coupled in a separate reaction vessel.

6. The process of claim 1, wherein the number of reaction vessels is less than the number of triplet codons to be randomized.

7. The process of claim 6, wherein two triplet codons to be randomized are coupled to a support in the same reaction vessel.

8. The process of claim 6, wherein the triplet codons are selected from the group consisting of TTT and GTT, TCT and CCT, TAT and CAT, TGT and CGT, CTG and ATG, CAG and GAG, ACT and GCT, AAT and GAT, TGG and GGG, and ATA and AAA.

9. A process of synthesizing oligonucleotides using individual monomers having at least one specified triplet codon at a predetermined position and the remaining positions having random triplet codons, said process comprising synthesizing the specified triplet codon on the mixed supports of claim 1 prior to dividing said mixed supports into separate reaction vessels.

10. The process of claim 9, wherein the first triplet codon is predetermined and is synthesized prior to step (1).

11. A process of synthesizing oligonucleotides having triplet codons which are diverse but biased toward a predetermined sequence, said process comprising:

(1) sequentially coupling monomers on a support to form a triplet codon with a predetermined sequence, said coupling being performed in a first reaction vessel;

(2) sequentially coupling monomers on a support to form a random triplet codon, said coupling being performed in a second reaction vessel;

(3) mixing the supports from said first and second reaction vessels; and

(4) dividing the mixed supports from step (3) and using the supports in step (1) and step (2), wherein steps (1) through (3) are repeated at least once.

12. The process of claim 11 further comprising (5) cleaving said oligonucleotide from said support.

13. The process of claim 11, wherein step (2) further comprises:

(2a) coupling a mixture of four different monomers for the first monomer of the random triplet;

(2b) coupling a mixture of four different monomers for the second monomer of the random triplet; and

(2c) coupling a pair of monomers selected from;

guanine and thymine monomers, or

cytosine and adenine monomers

for the third monomer of the random triplet.

14. The process of claim 13, wherein the four different monomers are adenine, guanine, cytosine and thymine.

15. The process of claim 13, wherein the four different monomers in steps (2a) and (2b) are present in equal proportions.

16. The process of claim 14, wherein the four different monomers in steps (2a) and (2b) are present in equal proportions.

17. The process of claim 13, wherein the four different monomers in steps (2a) and (2b) are present in unequal proportions.

18. The process of claim 14, wherein the four different monomers in steps (2a) and (2b) are present in unequal proportions.

19. The process of claim 13, wherein the guanine and thymine monomers of step (2c) are present in equal proportions.

20. The process of claim 13, wherein the guanine and thymine monomers of step (2c) are present in unequal proportions.

21. The process of claim 11, wherein the mixed supports from said first and second reaction vessels are divided into equal proportions.

22. The process of claim 11, wherein the mixed supports from said first and second reaction vessels are divided into unequal proportions.