Bead-Based Multiplexed Analytical Methods and Instrumentation

• US 20100130369A1
• Filed: 03/24/2008
• Published: 05/27/2010
• Est. Priority Date: 04/23/2007
• Status: Abandoned Application

• Current Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Original Assignee: Advanced Liquid Logic, Inc. (Illumina Incorporated)
• Inventors: Grichko, Varvara, Shenderov, Alexander, Kouri, Richard
• Application Number:
  US12/596,897
• Publication Number:
  US 20100130369A1
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2531/125 Rolling circle
• CPC Class Codes: C12Q 2563/155 Particles of a defined size, e.g. nanoparticles
• CPC Class Codes: C12Q 2563/179 the label being a nucleic acid

Abstract

Various methods, such as a method of detecting SNPs, involving (a) introducing onto a droplet actuator a solution comprising genomic DNA, extension oligos and suspension array beads; (b) dispensing on the droplet actuator one bead per droplet; (c) cleaving DNA from the bead in each droplet; (d) amplifying the cleaved DNA; (e) detecting SNP signals and barcode signals from the amplified DNA.

Claims

1. A method of detecting SNPs, the method comprising:

(a) introducing onto a droplet actuator a solution comprising genomic DNA, extension oligos and suspension array beads;

(b) dispensing on the droplet actuator one bead per droplet;

(c) cleaving DNA from the bead in each droplet;

(d) amplifying the cleaved DNA;

(e) detecting SNP signals and barcode signals from the amplified DNA.

2. A method of detecting the SNP signals and barcode signals, the method comprising:

(a) preparing suspension arrays on a preparative droplet microactuator;

(b) preparing genomic DNA in a single-tube reaction using beads from the suspension arrays;

(c) arraying individual SNP-beads on a second droplet microactuator;

(d) preparing individual beads for SNP identification and barcode reading on the second droplet actuator; and

(e) detecting SNP signals and barcode signals on the second droplet actuator.

3. A method of preparing a suspension array, the method comprising:

(a) introducing a bead suspension onto a droplet microctuator;

(b) dispensing on the droplet actuator;

(i) the bead suspension into an array of droplets;

(ii) droplets comprising barcode sequences;

(iii) droplets comprising probes and binding reagents;

(c) combining the droplets dispensed in (a) and performing the binding reaction;

(d) recombining the suspension array.

4. The method of claim 3 wherein one or more of the dispensing, combining, and recombining steps is conducted using droplet operations.