Customer Application (system support)
Increasing Throughput and Quality of Sequencing by Automation of MultiScreen* Filter Technology on the Biomek® FX
David S. Wexler, Cristina Melero, and Marcy Engelstein
AGY Therapeutics, Inc., South San Francisco, CA 94080
Millipore Corporation, 17 Cherry Hill Drive, Danvers, MA 01923
AGY Therapeutics, Inc.
Millipore Corporation –
www.millipore.com
Poster Abstract
As Genomic applications move towards the support of Drug Discovery technologies, the need for processing large numbers of samples in an automated fashion continues to be extremely important. Specifically, DNA sequencing technology is a very important tool for looking at the molecular basis of disease. In this poster we describe techniques which were successful in improving sequence quality and the automation of these processes. The purity of a DNA template contributes significantly to the overall quality of a sequence. When sequencing from PCR* products, we observe quantitative improvements in number of successful sequences and read length if we purify the PCR reactions on the MultiScreen -PCR plate prior to its use as a template. This procedure is automated for use on a Biomek FX liquid handling station. We compare the use of ethanol precipitation to Millipore's MultiScreen -SEQ filter plate for sequencing reaction cleanup. Several improvements were seen when using Millipore's vacuum filtration based assay versus centrifugal based ethanol precipitation when analyzed on the ABI 3700 DNA sequencer. These improvements include throughput, sequence quality and read length. Ease of automation is an attribute of this filtration-based assay. Samples are introduced, purified and recovered from the surface of the membrane using an automated liquid handler. All filtrate is directed to waste eliminating the need for manifold assembly/disassembly which is time consuming and difficult to automate. We have automated both MultiScreen384 assays on the Biomek FX using two manifolds simultaneously to meet our aggressive throughput needs. Two Plates of 384 PCR or SEQ can be processed in approx 45 minutes without intervention.
Features
- PCR method purifies two 384 plates in under 46 minutes.
- Sequencing methods purifies two 384 plates in under 42 minutes
- Washing tips enables the method to use only one box for each method.
- Minimal setup time required.
- No filtrate collection required, only top access to membrane necessary.
- Observed improvements in read length due to purity of samples.
* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffman-LaRoche, Ltd.
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