TECHNOLOGY/BUSINESS OPPORTUNITY

Microdroplet PCR and Sample Preparation Systems and Methods

Opportunity: Lawrence Livermore National Laboratory (LLNL), operated by the Lawrence Livermore National Security (LLNS), LLC under contract with the U.S. Department of Energy (DOE), is offering the opportunity to license and commercialize its Microdroplet PCR technology and related systems and methods for sample preparation.

Background: The first lab-on-chip system for picoliter droplet generation and PCR amplification with real-time fluorescence detection has performed PCR in isolated droplets at volumes 1,000,000 smaller than commercial real-time PCR instruments. The system has been demonstrated using both DNA and RNA based organisms. The system utilized a shearing T-junction in a silicon device to generate a stream of monodisperse picoliter droplets that were isolated from the microfluidic channel walls and each other by the oil-phase carrier. An off-chip valving system stopped the droplets on-chip, allowing them to be thermally cycled through the PCR protocol without droplet motion. With this system, a 10-pL droplet, encapsulating less than one copy of viral genomic DNA through Poisson statistics, showed realtime PCR amplification curves with a cycle threshold of_18, 20 cycles earlier than commercial instruments. This combination of the established real-time PCR assay with digital microfluidics is ideal for isolating single-copy nucleic acids in a complex environment.

Description: LLNL has developed unique methods and technologies relevant to microdroplet PCR for performing parallel nucleic acid sorting, amplification, and characterization. LLNL's technology is further described in publications:

"On-chip, Single-copy, Real-time Reverse-Transcription PCR in Isolated Picoliter Droplets"

http://pubs.acs.org/cgi-bin/article.cgi/ancham/2008/80/i06/pdf/ac800048k.pdf

"On-Chip, Real-time, Single-copy Polymerase Chain Reaction in Picoliter Droplets"

http://pubs.acs.org/cgi-bin/article.cgi/ancham/2007/79/i22/pdf/ac701809w.pdf

In addition, LLNL has developed a suite of technologies covering systems and methods for sample preparation. LLNL's inventions can be applied to sample preparation steps prior to analysis of nucleic acids (e.g. PCR, gel-electrophoresis, and DNA sequencing). Using a series of continuous flow, fluidic-based separation systems that include acoustic focusing, dielectrophoresis, isotachoporesis, and electrophoresis, viral DNA can be extracted from a clinical sample containing bacteria, proteins, viruses, and nucleic acids.

Advantages: These microfluidic systems can be used in a variety of applications for rapid, highly sensitive assays. The sample prep advances will lead to an automated system of sample preparation without the need for user interaction. This will improve throughput and recovery and lead to reliable and reproducible pretreatment for a broad range of samples.

Potential Applications:

Uses of the inventions include:

•1) Biomedical applications-tracing identifying, and monitoring

outbreaks of infectious diseases; automated processing,

amplification, and detection of host or microbial and viral DNA or

RNA for medical purposes

•2) Food and beverage safety

•3) Forensics

4) Biological pathogen detection-identifying, detecting and monitoring bio-threat agents that

contain nucleic acid signatures

Development Status:

LLNL has filed patent applications for the microdroplet PCR portfolio. LLNL is in the process of filing patent applications for the sample preparation portfolio.

LLNL is seeking industry partners with a demonstrated ability to bring such inventions to the market. Moving critical technology beyond the Laboratory to the commercial world helps our licensees gain a competitive edge in the marketplace. All licensing activities are conducted under policies relating to the strict nondisclosure of company proprietary information.

Please visit the IPO website at http://ipo.llnl.gov/workwithus/partneringprocess.php

for more information on working with LLNL and the industrial partnering and technology transfer process.

Note: THIS IS NOT A PROCUREMENT. Companies interested in commercializing LLNL's should provide a written statement of interest, which includes the following:

1. Company Name and address.

2. The name, address, and telephone number of a point of contact.

•3. A description of corporate expertise and facilities relevant to commercializing this technology.

Written responses should be directed to:

Lawrence Livermore National Laboratory

Industrial Partnerships Office

P.O. Box 808, L-795

Livermore, CA 94551-0808

Attention: FBO 173-08

Please provide your written statement within thirty (30) days from the date this announcement is published to ensure consideration of your interest in LLNL's Microdroplet PCR and Sample Preparation Systems and Methods.