Assays and other reactions involving droplets

Patent metadata

Field	Value
Publication number	US-9017948-B2
Application number	US-201414172266-A
Country	US
Kind code	B2
Filing date	Feb 4, 2014
Priority date	Mar 7, 2007
Publication date	Apr 28, 2015
Grant date	Apr 28, 2015

Abstract

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.

First claim

What is claimed is:

1. A method of amplification comprising:
a. generating a first plurality of droplets comprising polynucleotide primers, wherein the first plurality of droplets excludes polynucleotides to be amplified;
b. generating a second plurality of droplets comprising polynucleotides to be amplified, wherein the polynucleotides are obtained from a whole genome;
c. merging first droplets of the first plurality of droplets with second droplets of the second plurality of…

Assignees

• Harvard College

Inventors

• Agresti Jeremy
• Chu Liang-Yin
• Weitz David A
• Kim Jin-Woong
• Rowat Amy
• Sommer Morten
• Dantas Gautam
• Church George

Classifications

• G01N2015/1006

  Physics · mapped topic

• C12Q1/6834

  Chemistry & Metallurgy · mapped topic

• G01N15/1404

  Physics · mapped topic

• C12P19/34 Primary

  Chemistry & Metallurgy · mapped topic

• B01J13/0065

  Operations & Transport · mapped topic