Flexible Automated Platform for Blood Group Genotyping on DNA Microarrays Sandra Paris, Dominique Rigal, Valérie Barlet, Martine Verdier, Nicole Coudurier,

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Presentation transcript:

Flexible Automated Platform for Blood Group Genotyping on DNA Microarrays Sandra Paris, Dominique Rigal, Valérie Barlet, Martine Verdier, Nicole Coudurier, Pascal Bailly, Jean-Charles Brès The Journal of Molecular Diagnostics Volume 16, Issue 3, Pages 335-342 (May 2014) DOI: 10.1016/j.jmoldx.2014.02.001 Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Schematic representation of the workflow process for automated DNA-based blood group genotyping. Step 1: Collection of whole blood samples from donors for processing. Step 2: Genomic DNA extraction using a workstation that combined an automated liquid handling MagNA Starlet system (Hamilton Robotics) with the MagNA Pure 96 system (Roche Diagnostics). The MagNA Starlet system performs all pre-PCR processing, including sample distribution into processing plate and preparation of the PCR plate after nucleic acid extraction on the MagNA Pure system. Step 3: Multiplex PCR amplification on a Biometra thermal cycler. Inset: An electropherogram obtained after development of the multiplex PCR protocol. Step 4: Post-PCR processing performed on the Hamilton Starlet system (Hamilton Robotics). Hybridization of amplified products on the 96-well DNA microarrays is followed by washing. Step 5: Scanning of DNA microarrays on a fluorescence scanner LS200 (TECAN) to detect labeled multiplex PCR products. Inset: An image of a well obtained after array scanning. Step 6: After analysis of images using GenePix Pro software version 6.0 (Molecular Devices), genotypes assignment to each sample by comparing the calculated ratio for each SNP to the defined ratio limits. Phenotypes are predicted from genotypes. The Journal of Molecular Diagnostics 2014 16, 335-342DOI: (10.1016/j.jmoldx.2014.02.001) Copyright © 2014 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions