De novo synthesized gene libraries

Abstract

De novo synthesized large libraries of nucleic acids are provided herein with low error rates. Further, devices for the manufacturing of high-quality building blocks, such as oligonucleotides, are described herein. Longer nucleic acids can be synthesized in parallel using microfluidic assemblies. Further, methods herein allow for the fast construction of large libraries of long, high-quality genes. Devices for the manufacturing of large libraries of long and high-quality nucleic acids are further described herein.

1 - 20 . (canceled) 
     
     
         21 . A system for constructing a nucleic acid molecule comprising an identifier, the system comprising:
 (a) a first printhead configured to deposit a first component of the nucleic acid molecule comprising an identifier on a specified site on a substrate;   (b) a second printhead configured to deposit a second component of the nucleic acid molecule comprising an identifier on the specified site on the substrate; and   (c) a dispenser configured to deposit a reaction mix onto the specified site on the substrate to link the first component and the second component to construct the nucleic acid molecule,
 wherein the nucleic acid molecule comprising the identifier represents sequence information. 
   
     
     
         22 . The system of  claim 21 , wherein the dispenser comprises a third printhead configured to deposit the reaction mixture on the specified site on the substrate. 
     
     
         23 . The system of  claim 21 , wherein the first printhead, the second printhead, the dispenser, or a combination thereof comprise a plurality of nozzles. 
     
     
         24 . The system of  claim 21 , wherein the nucleic acid molecule comprises a dye-labeled nucleic acid molecule. 
     
     
         25 . The system of  claim 21 , wherein the first printhead, the second printhead, or the dispenser comprises a piezoelectric element or a thermoelectric element. 
     
     
         26 . The system of  claim 21 , wherein the reaction mixture comprises a ligase. 
     
     
         27 . The system of  claim 21 , wherein the substrate is configured for movement in the x-y plane, z-direction, or both. 
     
     
         28 . The system of  claim 21 , further comprising a vision system for aligning the first printhead, the second printhead, the dispenser, or any combination thereof with the substrate. 
     
     
         29 . The system of  claim 21 , further comprising a temperature control system, a vacuum device, or both. 
     
     
         30 . The system of  claim 29 , wherein the temperature control system comprises a thermal cycling device for performing nucleic acid amplification or assembly. 
     
     
         31 . The system of  claim 21 , wherein the first component, the second component, or both are building blocks, wherein the building blocks comprises about 1 to 50 nucleotides. 
     
     
         32 . The system of  claim 21 , wherein the substrate comprises a functionalization pattern. 
     
     
         33 . The system of  claim 21 , further comprising an alignment unit for aligning the substrate relative to the first printhead, the second printhead, or the dispenser. 
     
     
         34 . A method for constructing a nucleic acid molecule comprising an identifier, the method comprising:
 (a) depositing a first droplet comprising a first component of the nucleic acid molecule comprising an identifier on a specified site on a substrate using a first printhead;   (b) depositing a second droplet comprising a second component of the nucleic acid molecule comprising an identifier on the specified site on a substrate using a second printhead; and   (c) linking the first component and the second component to construct the nucleic acid molecule using a dispenser, wherein the linking is performed by dispending a reaction mix onto the specified site on the substrate, providing a condition necessary to link the first component and the second component, or both,
 wherein the nucleic acid molecule comprising the identifier represents sequence information. 
   
     
     
         35 . The method of  claim 34 , wherein linking the first component and the second component comprises using a ligase or click chemistry. 
     
     
         36 . The method of  claim 34 , further comprising incubating the specified site of the substrate. 
     
     
         37 . The method of  claim 34 , further comprising subjecting the specified site of the substrate to a given temperature, humidity, pressure, or gas content. 
     
     
         38 . The method of  claim 34 , further comprising capturing the nucleic acid molecule comprising the identifier. 
     
     
         39 . The method of  claim 34 , further comprising performing quality control of the nucleic acid molecule comprising the identifier. 
     
     
         40 . The method of  claim 34 , further comprising pooling the nucleic acid molecule with an identifier and at least one additional nucleic acid molecule.