Beckman Coulter Application (system and validated method support)
Automation of Amplification and Primer Extension Chemistries for Beckman Coulter’s GenomeLab™ SNPstream® Genotyping System
Keith Roby**, Laura Pajak**, Dana Campbell**, Zhiming Jiang**, Chad
Pittman** and Scott Boyer.
Beckman Coulter, Inc.
**Indianapolis, IN and ***Fullerton, CA
This presentation describes the utilization of the Biomek® FX Laboratory Automation Workstation for the process of SNP scoring using the SNPstream Genotyping System and assay reagents. Pre- and post-PCR* methods are executed on separate instruments to ensure that the overall hygience of the system is maintained thereby minimizing the possibility of cross-contamination.
Pre-PCR methods include Genomic DNA redistribution and multiplex reaction setup. Post-PCR methods include Exo/-SAP cleanup, multiplex single base primer extension and hybridization of the tagged extension products to the SNPstream assay plate.
The multiplex PCR and Primer extension protocols currently can accommodate up to 12 targets per reaction (i.e., a 1-plex reaction). Components required to generate amplicons in a multiplex reaction and to process these targets with SNPware® Reagent kits will be described in the talk; this includes:
• Description of the Biomek FX configurations utilized for pre- and post-PCR methods
• Description of important features of the methods used to generate and process samples
• Description of results obtained when using these methods
Using the methodology described in the talk, processing of 8 plates (> 36,000 SNPs) can be achieved in approximately 6 hours.
* 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.
