Microfluidic devices and methods for gene synthesis
Certain aspects of the present invention provide devices and methods for preparing oligonucleotides and for assembling nucleic acid molecules using microfluidic devices.
1. A microfluidic system comprising:
a continuous carrier phase fluid comprising droplets that are immiscible with the carrier phase fluid;
at least one main channel adapted to carry the carrier phase fluid, the channel comprising interior walls; and
a plurality of defined surface-capture spots forming a pattern on the channel interior walls, at least one of the plurality of defined surface-capture spots comprising oligonucleotides and a surface area modification having a hydrophilic surface property different than the interior walls, wherein the hydrophilic surface property allows droplet absorption and/or attachment, the plurality of defined surface-capture spots comprising oligonucleotides being configured to allow assembly of the oligonucleotides into a target polynucleotide sequence.
2. The microfluidic system of claim 1 further comprising an inlet module upstream of the main channel.
3. The microfluidic system of claim 1 further comprising an outlet module downstream of the main channel.
4. The microfluidic system of claim 1 wherein the surface modification affects the hydrophilic surface property of at least one of the plurality of defined surface-capture spots.
5. The microfluidic system of claim 1 wherein the carrier phase fluid is a water-immiscible fluid, and wherein the channel interior walls comprise an hydrophobic surface.
6. The microfluidic system of claim 1 wherein each of the plurality of defined surface-capture spots comprises oligonucleotides having a different predetermined sequence that together form the target polynucleotide sequence.
7. The microfluidic system of claim 6 wherein the plurality of defined surface-capture spots form a high density oligonucleotide array.
8. The microfluidic system of claim 1 further comprising a surface-capture module, wherein the surface capture module comprises the plurality of defined surface-capture spots and a force transducer.
9. The microfluidic system of claim 8 wherein the force transducer is an electric field, a magnetic field, a mechanical force, an optically induced force or any combination thereof.
10. The microfluidic system of claim 1 further comprising a force transducer for controlling 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 droplet is water-based and the carrier phase fluid is oil-based.
13. The microfluidic system of claim 1 , wherein one or more of the defined surface-capture spots is passivated.
14. The microfluidic system of claim 13 , wherein the passivated surface-capture spot is protected with a reagent.
15. The microfluidic system of claim 14 , wherein the reagent is oil or short chain hydrocarbon.
16. The microfluidic system of claim 1 , further comprising one or more reagents for performing amplification, assembly, and/or error correction, contained within the droplets.
17. A microfluidic system for assembling a target polynucleotide having a predetermined sequence comprising:
a continuous carrier phase fluid comprising droplets that are immiscible with the carrier phase fluid;
a plurality of channels adapted to carry the carrier phase fluid, the channel comprising hydrophobic interior walls;
a plurality of defined surface-capture spots on the interior walls, the surface-capture spots being chemically modified to confer hydrophilic surface property that allows droplet absorption and/or attachment, each defined surface-capture spot comprising oligonucleotides having a different predetermined subsequence that together form the predetermined sequence of the target polynucleotide; and
a force transducer for controlling droplet displacement from a first defined surface-capture spot to a second defined surface-capture spot to allow assembly of the oligonucleotides into the target polynucleotide.
18. The microfluidic system of claim 17 further comprising a reaction chamber for the assembly of the target polynucleotide.
19. The microfluidic system of claim 17 wherein the force transducer is an electric field, a magnetic field, a mechanical force, an optically induced force or any combination thereof.
20. The microfluidic system of claim 17 , further comprising one or more one or more reagents for performing amplification, assembly, and/or error correction, contained within the droplets.