Fluid processing device
View Patent ↗A fluid processing device and method of using the device are provided. The fluid processing device can include a substrate with a fluid processing pathway at least partially formed in or on the substrate. The fluid processing pathway can include an input end, at least one output end, a first input opening, a plurality of reaction sites each in fluid communication with the first input opening and arranged between the first input opening and the at least one output end. The fluid processing pathway can include a plurality of second input openings including two or more in fluid communication respectively with each of the reaction sites, the second input openings being arranged with the reaction site disposed between the at least one output end and the second input openings. The fluid processing device can include one or more output openings in fluid communication with one or more of the plurality of reaction sites and arranged at the at least one output end of the fluid processing pathway.
1. A method of synthesizing a polynucleotide sequence comprising:
providing a fluid processing device comprising a reaction chamber and first and second input openings in fluid communication with the reaction chamber;
introducing a protected first nucleotide monomer into the second input opening of a fluid processing device;
moving the protected first nucleotide monomer by capillary action from the second input opening into the reaction chamber;
attaching the protected first nucleotide monomer to a support disposed in the reaction chamber to form a protected first supported nucleotide monomer;
introducing a first deprotecting reagent into the first input opening;
moving the first deprotecting reagent by capillaiy action from the first input opening into the reaction chamber to form a deprotected first supported nucleotide monomer in the reaction chamber;
introducing a wash reagent into the first input opening;
moving the wash reagent by capillary action from the first input opening into the reaction chamber;
moving the first deprotecting reagent out of the reaction chamber;
introducing a protected second nucleotide monomer into the second input opening;
moving the protected second nucleotide monomer by capillary action from the second input opening into the reaction chamber; and
contacting the protected second nucleotide monomer with the deprotected first nucleotide monomer in the reaction chamber to form a supported polynucleotide sequence.
2. The method of claim 1 , wherein each of the protected first and protected second nucleotide monomers comprises a dimethyltrityl-protected phosphoramidite nucleotide monomer.
3. The method of claim 1 , further comprising:
introducing a wash reagent into the first input opening;
moving the wash reagent by capillary action from the first input opening into the reaction chamber; and
moving the wash reagent out of the reaction chamber.
4. The method of claim 3 , further comprising:
moving additional protected nucleotide monomer, deprotecting reagent, and wash reagent, into the reaction chamber to lengthen the supported polynucleotide sequence.
5. The method of claim 3 , further comprising:
moving a cleaving reagent by capillary action into the reaction chamber; and
cleaving the supported polynucleotide sequence from the support to form a cleaved polynucleotide sequence.
6. The method of claim 5 , further comprising removing the cleaved polynucleotide sequence from the reaction chamber.
7. The method of claim 1 , wherein the reaction chamber comprises a high surface area support material.
8. The method of claim 7 , wherein the high surface area support material comprises a removable particle, the supported polynucleotide sequence is attached to the removable particle, and the method further comprises removing the supported polynucleotide sequence, attached to the removable particle, from the reaction chamber.
9. The method of claim 7 , wherein the high surface area support material comprises a removable particle, the supported polynucleotide sequence is attached to the removable particle, and the method further comprises cleaving the supported polynucleotide sequence from the removable particle.
10. The method of claim 9 , wherein the cleaving occurs in the reaction chamber to form a cleaved polynucleotide sequence, and the method further comprises removing the cleaved polyruicleotide sequence from the reaction chamber.
11. The method of claim 9 , further comprising removing the supported polynucleotide sequence, attached to the removable particle, from the reaction chamber, wherein the cleaving occurs outside of the reaction chamber.
12. A method of synthesizing a polynucleotide sequence comprising:
providing a fluid processing device comprising a reaction chamber and first and second input openings in fluid communication with the reaction chamber;
introducing a protected first nucleotide monomer into the second input opening of a fluid processing device;
moving the protected first nucleotide monomer by centripetal force from the second input opening into the reaction chamber;
attaching the protected first nucleotide monomer to a support disposed in the reaction chamber to foim a protected first supported nucleotide monomer;
introducing a first deprotecting reagent into the first input opening;
moving the first deprotecting reagent by centripetal force from the first input opening Into the reaction chamber to form a deprotected first supported nucleotide monomer in the reaction chamber;
moving the first deprotecting reagent out of the reaction chamber;
introducing a first wash reagent into the first input opening;
moving the first wash reagent by centripetal force from the first input opening into the reaction chamber;
introducing a protected second nucleotide monomer into the second input opening;
moving the protected second nucleotide monomer by centripetal force from the second input opening into the reaction chamber;and
contacting the protected second nucleotide monomer with the deprotected first supported nucleotide monomer in the reaction chamber to form a supported polynucleotide sequence.
13. The method of claim 12 , wherein the step of moving the first deprotecting reagent out of the reaction chamber occurs after the step of introducing the first wash reagent into the first input opening.
14. The method of claim 12 , wherein the step of moving the first deprotecting reagent out of the reaction chamber occurs before the step of introducing the first wash reagent into the first input opening.
15. The method of claim 12 , wherein each of the protected first and protected second nucleotide monomers comprises a dimethyltrityl-protected phosphoramidite nucleotide monomer.
16. The method of claim 12 wherein the fluid processing device further comprises a valve disposed between the second input opening and the reaction chamber and which is capable of interrupting the fluid communication therebetween, and the method further comprises controlling the moving of the protected first nucleotide monomer from the second input opening into the reaction chamber by actuating the valve.
17. The method of claim 12 , further comprising:
moving a cleaving reagent by centripetal force into the reaction chamber; and
cleaving the supported polynucleotide sequence from the support to form a cleaved polynucleotide sequence.
18. The method of claim 17 , further comprising removing the cleaved polynucleotide sequence from the reaction chamber.
19. The method of claim 17 , wherein the fluid processing device further comprises an output chamber in fluid communication with the reaction chamber, and the method further comprises moving the cleaved polynucleotide sequence to the output chamber.
20. The method of claim 12 , wherein the reaction chamber comprises a high surface area support material.
21. The method of claim 20 , wherein the high surface area support material comprises a removable particle, the supported polynucleotide sequence is attached to the removable particle, and the method further comprises removing the supported polynucleotide sequence, attached to the removable particle, from the reaction chamber.
22. The method of claim 20 , wherein the high surface area support material comprises a removable particle, the supported polynucleotide sequence is attached to the removable particle, and the method further comprises cleaving the supported polynucleotide sequence from the removable particle.
23. The method of claim 22 , wherein the cleaving occurs in the reaction chamber to form a cleaved polynucleotide sequence, and the method further comprises removing the cleaved polynucleotide sequence from the reaction chamber.
24. The method of claim 22 , further comprising removing the supported polynucleotide sequence, attached to the removable particle, from the reaction chamber, wherein the cleaving occurs outside of the reaction chamber.
25. The method of claim 12 , further comprising:
introducing a second wash reagent into the first input opening;
moving the second wash reagent by centripetal force from the first input opening into the reaction chamber; and
moving the second wash reagent out of the reaction chamber.
26. The method of claim 25 , wherein the first and second wash reagents are the same.
27. The method of claim 25 wherein the fluid processing device further comprises a valve disposed between the first input opening and the reaction chamber, and capable of interrupting the fluid communication between the reaction chamber and the first input opening, and the method further comprises controlling the moving of the first wash reagent from the first input opening into the reaction chamber by actuating the valve.
28. The method of claim 25 , further comprising:
separately moving addition protected nucleotide monomer, deproecting reagent, and wash reagent, into the reaction chamber to lengthen the supported polynucleotide sequence.
29. The method of claim 28 , wherein the fluid processing device further comprises a waste chamber in fluid communication with the reaction chamber, and wherein moving the first deprotecting reagent out of the reaction chamber comprises moving the first deprotecting reagent from the reaction chamber to the waste chamber.