omation mbers - Society for Laboratory Automation and Screening

9:30 am Wednesday, February 4 Emerging Technologies – Cell Based Screening Room A3
Matthew Cook
TTP LabTech
Melbourn Science Park
Melbourn, Herts, SG8 6EE United Kingdom
matthew.cook@ttplabtech.com
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Co-Author(s)
Olivier Dery
BD Biosciences Clontech
Detection of BD Living Colors Novel Fluorescent Proteins Using the Acumen Explorer
With the sequencing of many genomes now completed, biologists are faced with the challenge of deciphering the
function and association of an immense number of resulting proteins. Understanding the specific processes and
actions of proteins and chemicals that comprise the cells and tissues of an organism, will be a necessary next step
to elucidating cellular function in healthy and disease states. Simultaneous and quantitative measurement of the
level of expression, for tens of thousands of genes aids in the definition of a comprehensive molecular phenotype
of cells and cellular processes. The Acumen Explorer exploits the power of fluorescence to monitor and
elucidate subtle changes in inter and intracellular biochemical events. Proprietary Novel Fluorescent Proteins (NFP)
from BD Biosciences Clontech include fluorescent proteins cloned from reef corals as well as from the jellyfish
Aequorea coerulescens. From this portfolio, five distinct proteins, AcGFP1 (monomer), ZsGreen1, ZsYellow1,
AsRed2 and DsRed2 are excited by a 488 nm Argon Ion laser. The resulting fluorescence emissions are directed to
four photo multiplier tubes. Three colors/wavelengths regions can be scanned simultaneously thereby allowing true
multiplexing with NFPs. The proprietary software algorithms permitted measurement in HEK293 cells of:
• Single and mixed populations of cells expressing NFP genes.
• Gene expression of two different colored NFP genes.
• Detection of proteins linked to different colored NFP localized to cell compartments.
The versatility of the Acumen Explorer in combination with BD Biosciences Clontech’s growing portfolio of NFPs
provides an excellent tool for the investigation of functional genomic applications.
9:45 am Wednesday, February 4 Emerging Technologies – Cell Based Screening Room A3
Johannes Dapprich
Generation Biotech, LLC
32 Pin Oak Drive
Lawrenceville, New Jersey 08648
jdapprich@generationbiotech.com
Co-Author(s)
Cynthia Turino, Sarah Jones, Colleen Murphy, Nancy Murphy
GenoVision, Inc.
Automated Molecular Haplotyping Through Physical Separation of DNA
Tine Thorbjornsen
Qiagen AS
Laurie Burdett, Marcello Fernandez-Vina
C.W. Bill Young Marrow Donor Program
We report the automation of a method, Haplotype-Specific Extraction (HSE), that allows the physical separation
of diploid genomic DNA into its haploid components. Magnetic beads are selectively attached to polymorphic
sites and used to isolate targeted, double-stranded DNA from a heterozygous mixture, thereby establishing
individual patient’s haplotypes within hours without knowledge of familial information. Automated Haploseparations
were carried out on several different liquid handling robots capable of handling 6, 48 and 96 samples
in different formats. This enables efficient, parallel and reliable processing of multiple samples. Haplo-separated
DNA is directly analyzed with kits and assays already in use for genotyping. The method was used on potential
organ donor samples to separate parts of chromosome 6 containing the HLA (Human Leukocyte Antigen)
locus. The highly polymorphic HLA-locus is routinely typed before transplantations can be carried out. Frequent
recombination events throughout the evolution of the locus have led to numerous allele pair combinations that, in
sum, lead to the same diploid genotype information as other allele pairs. HSE permits the unambiguous typing of
such allele pair combinations that may otherwise fail to be resolved by conventional sequence-based typing (SBT)
and sequence-specific oligonucleotide probes (SSOP). The purification of alleles allows the identification of known
or new haplotypes directly from genomic DNA.
PODIUM ABSTRACTS