LabAutomation 2006 - SLAS

TP55
Eric Shain
Abbott Molecular
Des Plaines, Illinois
eric.shain@abbott.com
Where Laboratory Technologies Emerge and Merge
Automation of Results Validity Checking on the m2000 Real Time PCR System
The Abbott m2000 system (not available in the U.S.) uses a novel, robust data reduction approach that allows analysis of real time PCR
signals and confirmation of results validity for the IVD market without need for user interaction or inspection. Historically, analysis of PCR
reactions has required visual inspection of the analyzed PCR growth curves to assure reliable results. The m2000 data reduction algorithm
utilizes both a threshold based analysis (Ct method), and a curve shape analysis (MaxRatio method) to analyze real time PCR signals. An
automatic baseliner handles transients in early cycles as well as early rising signals associated with high concentration samples. While the
Ct method provides linear and precise quantitation, it can be sensitive to anomalies in the baseline portion of the signal. The MaxRatio
method is an alternative algorithm for analyzing real time PCR signals that is highly robust to signal anomalies. It calculates quantitative
cycle numbers independent of Ct as well as a relative measure of reaction efficiency. These measures assure reliable reactive/non-reactive
determinations for a PCR reaction and also check the quality of the Ct value. In addition, curve shape analysis within the MaxRatio method
can provide an indication of reaction normality/abnormality. Result validity is evaluated on several levels including the integrity of the raw
fluorescence signals, amplification curve shape analysis, cycle number validity, internal control evaluation and plate controls.
TP56
Chris Bridge
DNA Research Innovations Ltd
Kent Science Park, United Kingdom
chris.bridge@invitrogen.com
Co-Author(s)
M, Crow
M.Baker
Nucleic Acid Purification From a Variety of Biological Samples Using ChargeSwitch ®
Technology in Coated Plate Format
In recent years there has been a drive towards higher and higher throughput in automated nucleic acid purification techniques. Here we
present a purification method specifically designed for integration into high throughput automation processes. ChargeSwitch ® chemistry
has been applied to 96 and 384 well microtitre plates to enable single-tube processing of a wide range of biological samples, offering fast,
efficient quantification-free purification directly into downstream applications such as PCR amplification or cycle sequencing.
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