SMALL SCALE DNA SYNTHESIS USING POLYMERIC SOLID SUPPORT WITH FUNCTIONALIZED REGIONS

• US 20010055761A1
• Filed: 10/29/1999
• Published: 12/27/2001
• Est. Priority Date: 10/29/1999
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies Europe BV (Agilent Technologies, Inc.)
• Inventors: KANEMOTO, ROY H., PERBOST, MICHAEL G. M.
• Application Number:
  US09/429,054
• Publication Number:
  US 20010055761A1
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: B01J 2219/00454 by chemical cleavage from the solid support
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00673 Slice arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• 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
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/14 Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support

Abstract

The invention relates to method and apparatus for synthesis of polymers, and specifically teaches the use of polymeric sheets suitable for the synthesis of small quantities of oligonucleotides such as DNA.

The polymeric sheets may be formed from a variety of materials, and wells or walled chambers formed by drilling or molding. The surfaces designated for synthesis product attachment sites are suitably functionalized. The well or walled chambers are of predetermined depth, corresponding to the relative amount of product desired to be synthesized per well. Sheets placed on an X-Y platform are amenable to automated synthesis protocols. After synthesis, collection of product may be immediate, or sheets may be stored and collection from predetermined wells may be made as desired. Many oligonuclotidess may be simultaneously synthesized, and quantities suitably controlled to ensure efficient production of desired product.

Claims

1. In a method for synthesizing oligonucleotides attached to a solid support, said method comprising sequentially coupling bases to form said oligonucleotide, wherein the improvement comprises sequentially coupling bases to dimensionally specific, functionalized regions within a polymeric support material.

2. In a method as in

claim 1

, further including the improvement of synthesized oligonucleotides contained within regions within a polymeric support material, where said support material may be stored and such synthesized oligonucleotides may be selectively collected by separating a portion of the support material from the remainder.

3. A method for oligonucleotide synthesis adapted for producing small quantities of oligonucleotide, comprising the steps:

Selecting a polymeric material in which there have been formed walled chambers of predetermined size;

Functionalizing said walled chambers sufficient to support oligonucleotide synthesis;

Synthesizing oligonucleotide within said walled chambers;

Harvesting said oligonucleotide from predetermined chambers.

4. A method as in

claim 3

, further comprising the steps of slicing the polymeric material into sheets of predetermined thickness and addressably orienting said sheets on an X-Y platform.

5. A method as in

claim 4

, further comprising the steps of delivering phosphoramidite solution into pre-selected chambers;

following the phosphoramidite reaction, the chambers are exposed to the next solution in the synthesis process, whether flush or reagent, repeating these two steps until the synthesis is complete.

6. A method as in

claim 3

wherein the harvesting is performed by introduction of a cleaving reagent into the chamber from which oligonucleotide is to be harvested.

7. A method as in

claim 3

wherein the polymeric material containing the oligonucleotide-containing chambers are stored for some time prior to harvesting.

8. A polymeric material adaptable for small scale oligonucleotide synthesis wherein said material is of predetermined dimensions and contains walled chambers suitable for creating functionalized regions for the synthesis of oligonucleotides.

9. A material as in

claim 8

wherein said material may be sliced to predetermined thickness in a predetermined orientation to the center of any chamber, such predetermination of thickness having the effect of selecting the amount of product that may be synthesized in such chamber by altering the wall space in any chamber.

10. A material as in

claim 8

wherein products of synthesis may be stored prior to collection, and wherein the material may be cut or otherwise divided into sub-sections for collection.