De novo synthesized gene libraries
Provided herein are devices for the manufacturing of high-quality oligonucleic acids. Longer nucleic acids, e.g., genes, can be synthesized in parallel using microfluidic assemblies described herein. Devices described herein include silicon plates having a plurality of channels in fluid communication with a plurality of microchannels. The number of microchannels and dimensions of the microchannels provide for rapid exchange of chemical exposure during de novo synthesis of oligonucleic acids.
1. A device for synthesizing oligonucleotides, the device comprising:
a silicon plate having formed therein:
a plurality of first channels, wherein each first channel extends vertically to a depth into the silicon plate from an opening on a top side of the silicon plate, and wherein each first channel has a width of 0.5 mm to 1.5 mm; and
for each first channel, 50 to 500 second channels in fluid communication with the first channel, wherein each second channel extends vertically to a height from an opening on a bottom side of the silicon plate to the first channel, wherein each second channel has a width of 1 um to 100 um, wherein a ratio of width to height of each second channel is from 1 to 0.1, wherein each second channel comprises a plurality of oligonucleotides bound to a first interior surface of said second channel, wherein each of the oligonucleotides comprises at least 80 nucleotides in length, and wherein the device provides for rapid exchange of chemical exposure during de novo synthesis of oligonucleotides, and
wherein the first interior surface of said second channel comprises a first set of molecules, wherein each molecule of the first set of molecules is an aminosilane or N-(3-triethoxvsilvlpropvD-4-hydroxybutvramide.
2. The device of claim 1 , wherein the aminosilane comprises 11-acetoxyundecyltriethoxysilane, n-decyltriethoxysilane, (3-aminopropyl)trimethoxysilane, or (3-aminopropyl)triethoxysilane.
3. The device of claim 1 , wherein each first channel comprises a second interior surface, wherein the second interior surface comprises a second set of molecules, wherein each molecule of the second set of molecules is a passive functionalization agent, bound to the second interior surface and lacks a reactive group capable of binding to a nucleoside phosphoramidite.
4. The device of claim 3 , wherein the second set of molecules comprises a fluorosilane.
5. The device of claim 4 , wherein the fluorosilane is (tridecafluorotetrahydrooctyl)-triethoxysilane.
6. The device of claim 1 , wherein the plurality of first channels comprises 2 to 250 first channels.
7. The device of claim 1 , wherein a total number of the second channels in the silicon plate is at least 15,000 second channels.
8. The device of claim 1 , wherein a total number of the second channels in the silicon plate is at least 30,000 second channels.
9. The device of claim 1 , wherein each first channel of the plurality of first channels has a width of 1 mm to 1.5 mm.
10. The device of claim 9 , wherein each first channel of the plurality of first channels has a width of 1.15 mm.
11. The device of claim 1 , wherein each of the 50 to 500 second channels has a width of 4 um to 70 um.
12. The device of claim 11 , wherein each of the 50 to 500 second channels has a width of about 60 um.
13. The device of claim 1 , wherein the first interior surface of each of the 50 to 500 second channels is coated with a layer of silicon dioxide.
14. The device of claim 1 , wherein:
the first interior surface comprises an aminosilane bound thereto; and
each second channel comprises a second interior surface, wherein the second interior surface comprises a fluorosilane bound thereto.
15. The device of claim 1 , wherein the ratio of width to height is 0.2.
16. The device of claim 1 , wherein the ratio of width to height is 0.1.
17. The device of claim 1 , wherein the ratio of width to height is 0.5.
18. The device of claim 1 , wherein the 50 to 500 second channels comprise 100 to 200 second channels.
19. The device of claim 1 , wherein each of the 50 to 500 second channels has a height of 10 um to 100 um.