omation mbers - Society for Laboratory Automation and Screening

4:30 pm Tuesday, February 3 Genomics – Analytical Room A1
Mark Shannon
Applied Biosystems
850 Lincoln Centre Drive
Foster City, California 94404
shannome@appliedbiosystems.com
Co-Author(s)
Kathryn Hunkapiller, Julie Blake, Maria Roque-Biewer, David Ruff
Applied Biosystems
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Shashi Amur, Shirley Zhu, Vicky Seyfert-Margolis
Immune Tolerance Network
Multiplex Preamplification of Assays-on-Demand Product Targets Prior to Quantitative
PCR Analysis of Gene Expression in Blood
Real-time quantitative PCR (qPCR) is fast becoming an important alternative approach to microarrays for
profiling the transcriptional states of cells and tissues. However, a requirement for several nanograms (ng) of
cDNA per assay often limits the scale of such gene expression studies. To overcome this potential obstacle to
large-scale expression studies, we have developed a method for preamplifying as many as 1000 Assays-on-
Demand product targets in a single reaction prior to conventional quantitative PCR (qPCR). The method is
highly reproducible, allows quantification of expression for hundreds of genes from as little as 0.01ng of cDNA per
assay as starting material, and is close to 100% efficient for the vast majority of targets. We applied this method
to expression profiling of 920 immune response related genes in T cells that were treated with phytohemagglutinin
(PHA). For roughly 20% of the genes, expression was detected only with multiplex preamplification. Of these
genes, approximately 60% were differentially expressed in response to treatment. This method is potentially
applicable to other situations where starting material is limited such as laser-capture microdissected tissues,
paraffin-embedded fixed tissues, and needle-biopsies.
8:00 am Wednesday, February 4 Genomics – Instrumentation Room A1
Steven Gordon
Brooks-PRI Automation
15 Elizabeth Drive
Chelmsford, Massachusetts 01824
Parallab Technology: Integrated Nanoliter Genomic Workstation
The Life Sciences Group of Brooks Automation, Inc. has developed a fully automated, integrated platform to
perform nanoliter volume reactions. As an example, our standard DNA cycle sequencing reaction is done in a
total volume of 500 nanoliters. A key element in this innovation is the proprietary Nano-Pipetter that incorporates
96 glass capillary tubes that process all samples in parallel. The bench top Parallab 350 exploits the ability to
aspirate small and accurate reagent volumes and subsequently completes all of the reaction and purification steps
within the 96 miniature glass syringes. Each reagent is aspirated into the glass vessels, mixed, thermal cycled
and purified before finally being dispensed into an output plate for analysis. After each sample set is complete,
the Nano-Pipetter is decontaminated and reused, greatly reducing the number and cost of consumables normally
associated with completing a similar sample set. For a genomics lab, the Parallab 350 is ideally suited for a
range of applications, including cycle sequencing, PCR, SNP analysis, genotyping and end point analysis and can
complete 1,800 samples per 24-hour day. We believe that this unique combination of attributes (nanoliter reaction
volume, reduced consumable use and complete automation) offers a revolutionary approach to help accelerate
discoveries in modern molecular biological laboratories.
PODIUM ABSTRACTS