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SpotQC Titration Experiment

SpotQC Titration Experiment Eight different organism-specifc microarrays were hybridized with SpotQC at six different concentrations to determine the most ideal dilution to use for quality control. Dilutions were as follows: 2 µM (SpotQC protocol) 1 µM .5 µM .25 µM .1 µM .01 µM.

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SpotQC Titration Experiment

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  1. SpotQC Titration Experiment • Eight different organism-specifc microarrays were hybridized with SpotQC at six different concentrations to determine the most ideal dilution to use for quality control. • Dilutions were as follows: • 2 µM (SpotQC protocol) • 1 µM • .5 µM • .25 µM • .1 µM • .01 µM

  2. SpotQC Titration Experiment • Tested 8 different organism print lots: • QAF01122005A A. fumigatus • QBA12012004A B. Anthracis • QLM11222004A L. monocytogenes • 3QSA12292004A S. aureus • QSM01042005A S. mutans • 2QSP11012004A S. pneumoniae • QVC12032004A V. cholera • QYP12102004A Y. pestis

  3. SpotQC Titration Experiment • Print plates for each print lot varied in age: • Date Created: • QAF01122005A 7/30/04 • QBA12012004A 3/31/04 • QLM11222004A 5/20/04 • 3QSA12292004A 12/22/04 • QSM01042005A 4/8/04 (refreshed 12/3/04) • 2QSP11012004A 4/8/04 • QVC12032004A 3/11/04 • QYP12102004A 3/11/04

  4. SpotQC Titration Experiment • Plates tested were 70mers or Amplicons: • QAF01122005A 70mer • QBA12012004A 70mer • QLM11222004A 70mer • 3QSA12292004A 70mer • QSM01042005A 70mer • 2QSP11012004A Amplicon • QVC12032004A 70mer • QYP12102004A 70mer

  5. SpotQC Titration Experiment • Slides were hybridized according to SpotQC protocol. • All slides were scanned on same scanner at PMT of 650. • Spotfinder was used to generate data from each tif image. • Log2 of signal intensities were calculated. • Box plots of data were created using Origin 7.5. • Data is visually represented as varying print lots across fixed concentrations as well as fixed print lots across varying concentrations.

  6. Each chart below graphs the concentration of SpotQC across 8 different organism print lots.

  7. QBA 0.5uM QBA 2uM QBA 1uM QBA 0.25uM QBA 0.1 QBA 0.01uM

  8. Each chart below graphs an organism print lot at different SpotQC concentrations.

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