Caliper Life Sciences, Inc.

Jill M Baker

Anne R Kopf-Sill

Andrea W Chow

Michael R Knapp

Michael A Spaid

J Wallace Parce

Theo Teofanov Nikiforov

Sansan Lin

Methods are provided for detecting low copy nucleic acids of interest in a sample. In one method, a sample comprising a nucleic acid of interest is aliquotted into a plurality of reaction mixtures, at least two of which are single-copy reaction mixtures. The reaction mixtures are subjected to one or more amplification reactions while flowing through a channel of a microfluidic device. At least one of the reaction mixtures is formulated in an aqueous phase of an emulsion comprising aqueous droplets suspended in an immiscible liquid. The nucleic acid of interest is present as a single copy in at least one aqueous droplet of the aqueous phase prior to performing the amplification reaction(s). Amplification is performed on the reaction mixture when it is formulated in the emulsion. The nucleic acid is continuously flowed during a plurality of steps of the method.

Methods of detecting low copy nucleic acids

Systems for differentiating the lengths of nucleic acids of interest in a sample are provided. The system includes a microfluidic device, a detector, and a software system. The microfluidic device includes an amplification microchannel or microchamber containing a reaction mixture under conditions that provide one or more amplicons of the nucleic acid of interest. The detector is integral with or proximal to the microfluidic device and is configured to detect the amplicons as one or more signals from a homogenous mixture. The software system interprets one or more coincidentally detected signals to indicate lengths of one or more individual nucleic acid molecules of interest, thereby differentiating the lengths of the nucleic acids of interest.

System for differentiating the lengths of nucleic acids of interest in a sample

Methods for determining nucleic acid fragmentation status are provided. A nucleic acid of interest in a reaction mixture is contacted with two or more different probes complementary to sites separated by a point of potential fragmentation. The probes each comprise a detectable marker. The nucleic acid of interest is flowed into a detection region, where two or more coincident detectable marker signals from the probes are detected. Fragmentation status of the nucleic acid of interest is determined, coincident detection of signals from two or more of the different probes indicating the nucleic acid of interest is not fragmented between the probes.

Determining nucleic acid fragmentation status by coincident detection of two labeled probes

Methods for chromatographically separating materials, including the separation of materials in kinase or phosphatase assays, in microfluidic devices under positive or negative fluid pressure. Devices and integrated systems for performing chromatographic separations are also provided.

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Redwood City, CA, United States of America

Jill M Baker