High throughput oligonucleotide synthesizer

• US 5,681,534 A
• Filed: 07/20/1995
• Issued: 10/28/1997
• Est. Priority Date: 07/20/1995
• Status: Expired due to Fees

• Current Assignee: Richard S. Neves
• Original Assignee: Richard S. Neves
• Inventors: Neves, Richard S.
• Primary Examiner(s): KIM, CHRISTOPHER Y
• Application Number:
  US08/504,959
• Time in Patent Office: 831 Days
• Field of Search: 422/131, 422/62, 422/81, 422/110, 435/91.1, 435/91.2, 435/91.21, 435/91.3, 435/91.31, 536/25.3, 935/16, 935/17, 935/88, 137/561 A, 285/131, 285/150, 285/155
• US Class Current: 422/131
• 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 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00306 Reactor vessels in a multiple arrangement
• CPC Class Codes: B01J 2219/00391 Rotary valves
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00722 Nucleotides
• 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: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: Y10T 137/85938 Non-valved flow dividers

Abstract

The present invention relates to a high-throughput oligonucleotide synthesizer for synthesizing multiple oligonucleotides comprising a pre-existing low throughput oligonucleotide synthesizer which has been integrated with an increased throughput expansion module in order to increase the throughput capacity of the low throughput synthesizer. More specifically, the present invention comprises (a) a pre-existing low-throughput oligonucleotide synthesizer having at least one and no more than eight reactor positions, (b) a plurality of valve manifolds each having a single inlet port and a plurality of outlet ports, (c) a plurality of fluidic connection means each having a first end in fluid connection with one of said reactor positions of the pre-existing low-throughput oligonucleotide synthesizer and a second end in fluidic connection with the inlet port of one of the plurality of valve manifolds and (d) a plurality of reactor positions corresponding in number to the number of valve manifold outlet ports, each of the plurality of reactor positions (d) having a single reactor inlet such that each of said reactor inlets is in fluid connection with a corresponding valve manifold outlet port. Each of the plurality of reactor positions (d) is configured to accept a pre-packaged reactor column, such as a pre-packaged reactor column comprising a substrate having a first 3'"'"' base covalently attached thereto. Multiple oligonucleotides are synthesized by the high-throughput oligonucleotide synthesizer of the present invention by the delivery of a stream of reagents from the pre-existing low throughput synthesizer to the plurality of reactor positions (d). In this manner, the high-throughput oligonucleotide synthesizer increases the throughput capacity of a conventional oligonucleotide low throughput synthesizer having the standard 2 or 4 reactor positions to an automated operation of up to 64 reactor positions, and typically from 12 to 24 reactor positions.

Claims

1. A high-throughput oligonucleotide synthesizer for synthesizing multiple oligonucleotides comprising:

(a) a pre-existing low-throughput oligonucleotide synthesizer having no more than eight reactor positions;

(b) a plurality of valve manifolds, each of said valve manifolds having (1) a single inlet port and (2) a plurality of outlet ports;

(c) a plurality of fluidic connection means, said plurality of fluidic connection means being equal to the number of reactor positions of said pre-existing low-throughput oligonucleotide synthesizer, each of said fluidic connection means having a first end in fluid connection with one of said reactor positions of said pre-existing low-throughput synthesizer and a second end in fluidic connection with one of said inlet ports of said plurality of valve manifolds, such that each of said plurality of valve manifolds is in fluidic connection with one of said reactor positions of said pre-existing low-throughput oligonucleotide synthesizer; and

(d) a plurality of reactor positions corresponding in number to said plurality of outlet ports of said plurality of valve manifolds, each of said plurality of reactor positions comprising a single reactor inlet and a single reactor outlet and being capable of receiving a reactor column, such that each reactor inlet of said plurality of reactor positions is in fluidic connection with a corresponding valve manifold outlet port;

whereby the throughput capacity of the pre-existing low-throughput oligonucleotide synthesizer is multiplied by the number of plurality of outlet ports of said plurality of valve manifolds.

2. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein each of said plurality of fluidic connection means is in the form of an interconnect comprising a coupling unit having a first end attached to the inlet of said reactor position of said pre-existing low throughput synthesizer and having a second end connected by tubing means to said inlet port of one of said plurality of valve manifolds, said interconnect being configured in such a manner as to produce a zero dead volume attachment.

3. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein each of said plurality of valve manifolds comprises six outlet ports.

4. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein said pre-existing low-throughput oligonucleotide synthesizer comprises from two to four reactor positions.

5. The high-throughput oligonucleotide synthesizer in accordance with claim 4, wherein each of said plurality of valve manifolds comprises at least four outlet ports.

6. The high-throughput oligonucleotide synthesizer in accordance with claim 1, further comprising (e) a waste reservoir for receiving discharged effluent from said reactor outlets of said reactor positions (d).

7. The high-throughput oligonucleotide synthesizer in accordance with claim 1, further comprising (f) means for interfacing to a personal computer.

8. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein each of said plurality of valve manifolds has a linear configuration.

9. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein each of said plurality of valve manifolds has a radial configuration.

10. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein each of said plurality of reactor positions (d) is configured to accept a pre-packaged reactor column.

11. The high-throughput oligonucleotide synthesizer in accordance with claim 10, wherein said pre-packaged reactor column comprises a substrate having a first 3'"'"' base covalently attached thereto.

12. The high-throughput oligonucleotide synthesizer in accordance with claim 1, wherein said multiple oligonucleotides are synthesized by the delivery of a stream of reagents from said reactor position of said low-throughput oligonucleotide synthesizer to said reactor inlet of each of said plurality of said reactor positions (d).

13. The high-throughput oligonucleotide synthesizer in accordance with claim 12, wherein each of said reactor positions (d) contains a pre-packaged reactor column.

14. A high-throughput oligonucleotide synthesizer for synthesizing multiple oligonucleotides comprising:

(a) a pre-existing low-throughput oligonucleotide synthesizer having two reactor positions;

(b) two radial valve manifolds, each of said valve manifolds having (1) a single inlet port and (2) a plurality of outlet ports;

(c) two fluidic connection means, each of said fluidic connection means having a first end in fluid connection with one of said reactor positions of said pre-existing low-throughput synthesizer and a second end in fluidic connection with one of said inlet ports of said valve manifolds, such that each valve manifold is in fluidic connection with one of said reactor positions of said pre-existing low-throughput oligonucleotide synthesizer; and

(d) a plurality of reactor positions corresponding in number to said plurality of outlet ports of said two valve manifolds, each of said plurality of reactor positions comprising a single reactor inlet and a single reactor outlet and being capable of receiving a reactor column, such that each reactor inlet of said plurality of reactor positions is in fluidic connection with a corresponding valve manifold outlet port;

whereby the throughput capacity of the pre-existing low-throughput oligonucleotide synthesizer is multiplied by the number of plurality of outlet ports of said plurality of valve manifolds.

15. The high-throughput oligonucleotide synthesizer in accordance with claim 14, wherein each of said fluidic connection means is in the form of an interconnect comprising a coupling unit having a first end attached to the inlet of said reactor position of said pre-existing low throughput synthesizer and having a second end connected by tubing means to said inlet port of one of said of valve manifolds, said interconnect being configured in such a manner as to produce a zero dead volume attachment.

16. The high-throughput oligonucleotide synthesizer in accordance with claim 15, wherein each valve manifold comprises six outlet ports, each of said outlet ports being in fluidic connection with a corresponding reactor inlet of said plurality of reactor positions (d).

17. The high-throughput oligonucleotide synthesizer in accordance with claim 14, wherein each of said plurality of reactor positions (d) is configured to accept a pre-packaged reactor column comprising a substrate having a first 3'"'"' base covalently attached thereto, such that multiple oligonucleotides are synthesized by the delivery of a stream of reagents from each of the reactor inlets positions of said low throughput synthesizer to each of said reactor inlets of said plurality of reactor positions (d).