CREATION OF LIBRARIES OF DROPLETS AND RELATED SPECIES

• US 20150336068A1
• Filed: 07/29/2015
• Published: 11/26/2015
• Est. Priority Date: 09/19/2008
• Status: Abandoned Application

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Weitz, David A., Agresti, Jeremy
• Application Number:
  US14/812,930
• Publication Number:
  US 20150336068A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: B01J 13/04 by physical processes, e.g. drying, spraying
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00599 Solution-phase processes
• CPC Class Codes: B01J 2219/00709 Type of synthesis
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00716 Heat activated synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes

Abstract

The present invention is generally related to systems and methods for producing a plurality of droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, the fluidic droplets may be rigidified to form rigidified droplets (e.g., gel droplets). In certain embodiments, the droplets may undergo a phase change (e.g., from rigidified droplets to fluidized droplets), as discussed more herein. In some cases, a species may be added internally to a droplet by exposing the droplet to a fluid comprising a plurality of species.

Claims

1. A method, comprising:

providing a solution comprising;

a fluid;

a suspension of a plurality of rigidified species-containing droplets; and

a plurality of target analyte molecules; and

dispersing the solution into a plurality of microfluidic droplets, wherein at least a portion of the microfluidic droplets comprise a rigidified species-containing droplet and a target analyte molecule,

wherein the rigidified droplets are capable of fluidization.

2. The method of claim 1, wherein the dispersing comprises using microfluidic techniques.

3. The method of claim 1, wherein the dispersing comprises flowing the solution into a microfluidic channel comprising a second fluid.

4. The method of claim 3, wherein the second fluid is an immiscible fluid.

5. The method of claim 3, wherein the immiscible fluid is an oil.

6. The method of claim 1, wherein the rigidified species-containing droplets comprise a first group and a second group of rigidified droplets, wherein rigidified droplets in the first group comprise a first species, and wherein rigidified droplets in the second group comprise a second species distinguishable from the first species.

7. The method of claim 1, further comprising the step of fluidizing rigidified droplets in the microfluidic droplets.

8. The method of claim 1, wherein the species and the target analyte molecules interact.

9. The method of claim 6, wherein each group of rigidified droplets comprises at least one distinguishable identification element.

10. The method of claim 7, wherein the rigidified droplets are fluidized by exposure to a change in temperature.

11. The method of claim 7, wherein the rigidified droplets are fluidized by exposure to electromagnetic radiation.

12. The method of claim 7, wherein the electromagnetic radiation is ultraviolet light.

13. The method of claim 7, wherein the rigidified droplets are fluidized by exposure to a change in pH.

14. The method of claim 1, wherein the plurality of rigidified droplets comprise a polymer.

15. The method of claim 14, wherein the polymer comprises a plurality of crosslinks.

16. The method of claim 15, wherein the rigidified droplets are fluidized by causing the polymer crosslinks to cleave.

17. The method of claim 1, wherein the plurality of rigidified droplets comprise a gel.

18. The method of claim 17, wherein the wherein the rigidified droplets are caused to fluidize by causing the gel to reform a gel precursor.

19. The method of claim 1, wherein the target analyte molecules are nucleic acids.

20. The method of claim 19, further comprising the step of determining the sequence of the nucleic acids.

21. The method of claim 6, wherein at least about 5 groups of rigidified droplets are provided.

22. The method of claim 6, wherein at least about 10 groups of rigidified droplets are provided.

23. The method of claim 6, wherein at least about 20 groups of rigidified droplets are provided.

24. The method of claim 6, wherein at least about 64 groups of rigidified droplets are provided.

25. The method of claim 6, wherein at least about 100 groups of rigidified droplets are provided.

26. The method of claim 6, wherein at least about 4096 groups of rigidified droplets are provided.

27. The method of claim 6, wherein at least about 10,000 groups of rigidified droplets are provided.

28. The method of claim 6, wherein the distinguishable species are nucleic acids.