LabAutomation 2006 - SLAS

Where Laboratory Technologies Emerge and Merge
3:30 pm Monday, January 23, 2006 Track 3: High-Throughput Technologies Room: Learning Center
Wyndham Palm Springs Hotel
Mitchell Mutz
Co-Author(s)
ISunnyvale, California
Burt Kong
mutz@labcyte.com
Richard Ellson
Glen Krueger
Lawrence Lee
Michael Miller
A. Mark Bramwell
Self-Contained Environmental Lids for HTS Compound Preservation
Reduction of solvent hydration and evaporation is essential to ensure compound library integrity. HTS processes expose valuable library
compounds to the surrounding atmosphere during dispensing and some measurement procedures, giving an opportunity for moisture
to enter or for DMSO to escape from the sample container. Current efforts to limit the degradation of the compound with a controlled
environment are large scale, and often involve chambers filled with dry or inert gas. These chambers must be large enough for numerous
microplates, plate handling equipment and instruments. This paper describes a novel alternative to the macro-scale environmental
chamber. We miniaturize environmental control to the microplate scale with an “environmental lid” that provides a dry, DMSO-vapor
generating enclosure to protect the compound samples from both hydration and evaporation. Gravity secures the lid, and the lid is
compatible with standard plates and lid-handling automation. Short-term, “on the screening deck” applications such as preventing
evaporation of droplets dispensed into dry wells and reducing hydration in compound library plates are explored. Also, long-term
applications of the environmental lid for compound storage are presented.
4:00 pm Monday, January 23, 2006 Track 3: High-Throughput Technologies Room: Learning Center
Wyndham Palm Springs Hotel
Kurt Evans
Co-Author(s)
Ambion
Roy C. Willis
Austin, Texas
Quoc Hoang
kevans@ambion.com
Angela Burrel
Weiwei Xu
Sharmili Moturi
Mangkey Bounpheng
Xingwang Fang
Ambion
F I N A L I S T
High-Throughput Sample Preparation From Whole Blood for Gene Expression Analysis
Whole blood is an attractive sample source for assays involving nucleic acids because it is readily available, easily accessible, and rich in
genetic information. However, up to 70% of the mRNA (by mass) in whole blood total RNA is globin. Dominant globin mRNA comprises the
linear amplification during microarray sample preparation. Array analysis using total RNA containing a high percentage of globin mRNA has
been shown to decrease present calls, decrease call concordance and increase signal variation. Therefore, for gene expression analysis,
whole blood is typically fractionated before total RNA isolation.
We have developed a high throughput sample preparation technology that streamlines total RNA isolation from whole blood, globin mRNA
removal, and mRNA amplification and labeling for expression analysis. Our high-throughput blood RNA isolation method eliminates the
need for pre-fractionation of whole blood, thus minimizes the expression profile change during sample handling. Following RNA isolation,
we utilize a novel, non-enzymatic technology to rapidly deplete the alpha and beta globin mRNA, and the remaining RNA in the sample is
amplified using Ambion’s high-throughput amplification system, MessageAmp II-96.
Quantitative RTPCR showed our method effectively removed up to 95% of the globin mRNA from the isolated RNA while retaining the
normal levels of other mRNAs. When analyzed on Affymetrix gene chip, samples prepared with our method increased percent present
calls, decreased 3’/5’ ratios, and decreased sample to sample variability as compared to total RNA isolated from whole blood. Our method
allows quick and accurate expression analysis of relatively high numbers of blood samples.
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