Microfluidic devices and methods for gene synthesis

Patent metadata

Field	Value
Publication number	US-9968902-B2
Application number	US-201514946811-A
Country	US
Kind code	B2
Filing date	Nov 20, 2015
Priority date	Nov 25, 2009
Publication date	May 15, 2018
Grant date	May 15, 2018

Abstract

Certain aspects of the present invention provide devices and methods for preparing oligonucleotides and for assembling nucleic acid molecules using microfluidic devices.

First claim

The invention claimed is:

1. A microfluidic system comprising:
a continuous carrier phase fluid comprising at least one droplet in fluid communication with a solid support, wherein the at least one droplet is immiscible with the carrier phase fluid and wherein the at least one droplet comprises at least one input oligonucleotide, wherein the input oligonucleotide has sequence complementarity or overlaps with at least one immobilized oligonucleotide;
a plurality of defined surface-capture spots forming a pattern on the solid support,
wherein at least one of the plurality of defined surface-capture spots comprises a plurality of immobilized oligonucleotides and a patch having a hydrophilic surface property different than the solid support wherein the hydrophilic surface property allows droplet absorption and/or attachment; and
wherein the plurality of defined surface-capture spots comprising immobilized oligonucleotides being configured to allow assembly of the input oligonucleotides into a target polynucleotide sequence.

2. The microfluidic system of claim 1 , further comprising at least one main channel adapted to carry the carrier phase fluid.

3. The microfluidic system of claim 2 , wherein the channel comprises an interior wall.

4. The microfluidic system of claim 3 , wherein the plurality of defined surface-capture spots form a pattern on the interior wall.

5. The microfluidic system of claim 3 , wherein at least one defined surface-capture spot comprises a patch having a hydrophilic surface property different than the interior wall, wherein the hydrophilic surface property allows droplet absorption and/or attachment.

6. The microfluidic system of claim 2 , further comprising an inlet module upstream of the main channel and/or an outlet module downstream of the main channel.

7. The microfluidic system of claim 1 , wherein the carrier phase fluid is water immiscible.

8. The microfluidic system of claim 7 , wherein the solid support comprises a hydrophobic surface and wherein the defined surface-capture spots are hydrophilic.

9. The microfluidic system of claim 1 , wherein the solid support is a high density oligonucleotide array.

10. The microfluidic system of claim 1 , further comprising a force transducer that controls droplet displacement from a first defined surface-capture spot to a second defined surface-capture spot.

11. The microfluidic system of claim 10 , wherein the force transducer is an electric field, a magnetic field, a mechanical force, an optically induced force or any combination thereof.

12. The microfluidic system of claim 1 , wherein the at least one droplet is water-based and the carrier phase fluid is oil-based.

13. The microfluidic system of claim 1 , further comprising one or more reagents for performing amplification, assembly, and/or error correction, contained within the at least one droplet.

14. A microfluidic system for assembling a target polynucleotide having a predetermined sequence, comprising:
a continuous carrier phase fluid comprising at least one droplet in fluid communication with a solid support, wherein the at least one droplet is immiscible with the carrier phase fluid and comprises at least one input oligonucleotide;
a plurality of defined surface-capture spots forming a pattern wherein the solid support comprises immobilized oligonucleotides that together comprise a target polynucleotide and wherein the input oligonucleotide has sequence complementarity or overlaps with at least one immobilized oligonucleotide; and
a force transducer for controlling droplet displacement from a first surface-capture spot to a second surface-capture spot to allow assembly of the input oligonucleotides into the target polynucleotide.

15. The microfluidic system of claim 14 , wherein at least one defined surface-capture spots forming a pattern comprises a patch having a hydrophilic surface property different than the solid support, wherein the hydrophilic surface property allows droplet absorption and/or attachment.

16. The microfluidic system of claim 14 , wherein the solid support is a high density oligonucleotide array.

17. The microfluidic system of claim 14 , wherein the force transducer is an electric field, a magnetic field, a mechanical force, an optically induced force or any combination thereof.

18. The microfluidic system of claim 14 , further comprising one or more one or more reagents for performing amplification, assembly, and/or error correction, contained within the droplets.

Assignees

• Gen9 Inc

Inventors

• Chu Larry Li-Yang

Classifications

• B01J2219/0052

  in the shape of elongated tubes · CPC title

• B01J2219/00655

  the compounds being bound to magnets embedded in or on the solid supports · CPC title

• B01J2219/00286

  Reactor vessels with top and bottom openings · CPC title

• B01L2400/0478

  pistons · CPC title

• B01J2219/00722

  Nucleotides · CPC title