Microfluidic devices

Patent metadata

Field	Value
Publication number	US-12337287-B2
Application number	US-201715480739-A
Country	US
Kind code	B2
Filing date	Apr 6, 2017
Priority date	May 11, 2006
Publication date	Jun 24, 2025
Grant date	Jun 24, 2025

Abstract

The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.

First claim

What is claimed is:

1. A method for droplet formation for sample preparation for sequencing, the method comprising:
providing, via at least one inlet channel on a microfluidic substrate, an aqueous sample fluid comprising nucleic acid and at least one primer;
providing an immiscible first carrier fluid in a main channel on the microfluidic substrate;
producing a plurality of droplets comprising the aqueous sample fluid at a junction between the main channel and the at least one inlet channel;
copying at least a portion of the nucleic acid within one droplet of the plurality of droplets by extending the at least one primer to generate an extension product; and
breaking the plurality of droplets by introducing a droplet-destabilizing second fluid comprising a destabilizing surfactant, thereby recovering the aqueous sample fluid including the extension product from the plurality of droplets.

2. The method of claim 1 , wherein the aqueous sample fluid comprises one or more cells.

3. The method of claim 1 , further comprising subjecting the extension product to an amplification reaction.

4. The method of claim 3 , wherein the amplification reaction is a PCR reaction.

5. The method of claim 1 , wherein at least one oil supply line provides a constant flow of the first carrier fluid into a nozzle while the plurality of droplets are formed.

6. The method of claim 1 , wherein the first carrier fluid comprises an oil.

7. The method of claim 6 , wherein the oil comprises a fluorocarbon oil or a silicone oil.

8. The method of claim 6 , wherein the oil comprises a surfactant.

9. The method of claim 8 , wherein the surfactant is a fluorosurfactant.

10. The method of claim 1 , wherein the microfluidic substrate is a microfluidic chip.

11. The method of claim 1 , wherein the aqueous sample fluid comprises one or more beads.

12. The method of claim 1 , further comprising centrifuging the plurality of droplets and the droplet-destabilizing second fluid to promote complete coalescence of the plurality of droplets.

13. The method of claim 1 , wherein the destabilizing surfactant comprises a perfluorinated alcohol.

14. The method of claim 13 , wherein the perfluorinated alcohol comprises 1H,1H,2H,2H-Perfluoro-1-octanol.

15. The method of claim 1 , wherein the copying step uses a reverse transcriptase to produce complementary DNA.

16. A sample preparation method comprising:
flowing, via an inlet channel on a microfluidic substrate, an aqueous sample fluid comprising nucleic acid to a junction between the inlet channel and a main channel having an immiscible carrier fluid therein;
producing a plurality of droplets comprising the aqueous sample fluid surrounded by the immiscible carrier fluid in the main channel;
copying at least a portion of the nucleic acid within one droplet of the plurality of droplets by extending a primer to generate an extension product; and
recovering the aqueous sample fluid including the extension product from the plurality of droplets by introducing a droplet-destabilizing second fluid comprising a perfluorinated alcohol.

17. The method of claim 16 , wherein the perfluorinated alcohol comprises 1H,1H,2H,2H-Perfluoro-1-octanol.

18. The method of claim 16 , wherein the copying step uses a reverse transcriptase to produce complementary DNA.

19. The method of claim 16 , wherein the aqueous fluid sample comprises one or more cells.

20. The method of claim 16 , wherein the aqueous fluid sample comprises one or more beads.

Assignees

• Bio Rad Laboratories Inc

Inventors

• Link Darren Roy
• Weiner Michael
• Marran David
• Rothberg Jonathan M

Classifications

• C12Q2565/629

  being a microfluidic device · CPC title

• C12Q1/6855

  Ligating adaptors · CPC title

• C12N15/1068

  Template (nucleic acid) mediated chemical library synthesis, e.g. chemical and enzymatical DNA-templated organic molecule synthesis, libraries prepared by non ribosomal polypeptide synthesis [NRPS], DNA/RNA-polymerase mediated polypeptide synthesis · CPC title

• G01N27/3275

  Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title