Reagent storage on a droplet actuator

• US 9,631,244 B2
• Filed: 06/26/2012
• Issued: 04/25/2017
• Est. Priority Date: 10/17/2007
• Status: Active Grant

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Foley, Jennifer, Burde, Stefan, Pamula, Vamsee K., Pollack, Michael G.
• Primary Examiner(s): Andres, Janet L
• Assistant Examiner(s): Chestnut, Barry A
• Application Number:
  US14/131,061
• Publication Number:
  US 20140141409A1
• Time in Patent Office: 1,764 Days
• CPC Class Codes: B01L 2200/0678 Facilitating or initiating evaporation
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2400/0427 Electrowetting
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 3/502792 for moving individual droplets on a plate, e.g. by locally altering surface tension
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/703 Viruses associated with AIDS
• CPC Class Codes: C12Q 2523/308 Adsorption or desorption
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2531/113 PCR
• CPC Class Codes: C12Q 2563/159 Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode

Abstract

A method of providing a droplet comprising one or more reagents, the method comprising, depositing a first aqueous droplet comprising the one or more reagents on a surface; drying the droplet to yield a dried composition on the surface comprising the one or more reagents; covering the dried composition with oil; and causing a second aqueous droplet in the oil to contact the dried composition and thereby resuspend one or more reagents.

Claims

1. A method of providing a droplet comprising one or more reagents, the method comprising:

(a) depositing a first aqueous droplet comprising the one or more reagents on a surface;

(b) drying the first aqueous droplet to yield a dried composition on the surface comprising the one or more reagents;

(c) covering the dried composition with oil; and

(d) causing a second aqueous droplet in the oil to contact the dried composition by pulsing or shuttling the second aqueous droplet and thereby resuspending one or more reagents.

2. The method of claim 1, wherein the first aqueous droplet further comprises a stabilizing agent.

3. The method of claim 2, wherein the stabilizing agent comprises a sugar.

4. The method of claim 3, wherein the sugar is selected from a group consisting of:

dextrans, sucroses, and trehaloses.

5. The method of claim 2, wherein the stabilizing agent comprises a polymer.

6. The method of claim 2, wherein the one or more reagents comprise a reagent selected from a group consisting of:

beads, proteins, nucleic acids, salts, sugars, and surfactants.

7. The method of claim 1, wherein the one or more reagents comprise reagents selected for amplifying a target nucleic acid when combined with a sample comprising the target nucleic acid.

8. The method of claim 1, wherein the one or more reagents comprises an antibody.

9. The method of claim 1, wherein the one or more reagents comprises an antibody attached to a bead.

10. The method of claim 1, wherein the one or more reagents comprises a protease.

11. The method of claim 1, wherein the one or more reagents comprises protease K.

12. The method of claim 1, wherein the one or more reagents comprises a lectin.

13. The method of claim 1, wherein the one or more reagents comprises a phaseoulus vulgaris agglutinin.

14. The method of claim 1, wherein the one or more reagents comprises a bead.

15. The method of claim 1, wherein the one or more reagents comprises a virus.

16. The method of claim 1, wherein the one or more reagents comprises a spore.

17. The method of claim 1, wherein the one or more reagents comprises a bacteria.

18. The method of claim 1, wherein the one or more reagents comprises a fungus.

19. The method of claim 1, wherein the one or more reagents comprises an armored RNA.

20. The method of claim 1, wherein the one or more reagents comprises an armored DNA.

21. The method of claim 1, wherein the one or more reagents comprises a bacteriophage.

22. The method of claim 1, wherein the one or more reagents comprises MS2.

23. The method of claim 1, wherein the one or more reagents comprises a polymer.

24. The method of claim 1, wherein the one or more reagents comprises a temperature sensitive polymer.

25. The method of claim 1, wherein the one or more reagents comprises a fluorophore.

26. The method of claim 1, wherein the one or more reagents comprises a nucleic acid.

27. The method of claim 1, wherein the one or more reagents comprises a lysis reagent.

28. The method of claim 1, wherein the one or more reagents comprises a buffer.

29. The method of claim 1, wherein the one or more reagents comprises a magnetically responsive bead.

30. The method of claim 1, wherein the second aqueous droplet has a volume ranging from 10 picoliters to 10 milliliters.

31. The method of claim 1, wherein the first aqueous droplet has a volume ranging from one nanoliter to 3 milliliters.

32. The method of claim 1, wherein the first aqueous droplet has a volume ranging from five nanoliters to one milliliter.

33. The method of claim 1, wherein the second aqueous droplet has a volume ranging from 10 picoliters to 10 milliliters.

34. The method of claim 1, wherein the second aqueous droplet has a volume ranging from 100 picoliters to 5 milliliters.

35. The method of claim 1, wherein the second aqueous droplet has a volume ranging from 50 nanoliters to 2 milliliters.

36. The method of claim 1, wherein the second aqueous droplet has a volume ranging from 100 nanoliters to 0.5 milliliters.

37. The method of claim 1, wherein the first aqueous droplet comprises a surfactant.

38. The method of claim 1, wherein the second aqueous droplet comprises a surfactant.

39. The method of claim 1, wherein the surface comprises an electrode.

40. The method of claim 1, wherein the surface comprises a plastic surface.

41. The method of claim 1, wherein the surface is hydrophobic.

42. The method of claim 1, wherein the surface comprises an electrode and the dried composition is positioned on the electrode.

43. The method of claim 42, wherein the electrode is a component of an array of electrodes.

44. The method of claim 43, wherein the array of electrodes is configured to conduct droplet operations on the surface.