1 / 22

Determining Optimal Dilution of SpotQC in Titration Experiment with Microarrays

This experiment investigated the ideal dilution of SpotQC using eight distinct organism-specific microarrays, hybridized at six different concentrations (2 µM, 1 µM, 0.5 µM, 25 µM, 1 µM, and 0.1 µM). The microarrays tested included strains like A. fumigatus, B. anthracis, L. monocytogenes, and others. All slides were scanned under consistent conditions, and Log2 signal intensities were computed using Spotfinder. Data visualization included box plots depicting variations in signal intensities across different concentrations and print lots to establish quality control standards.

fritz
Télécharger la présentation

Determining Optimal Dilution of SpotQC in Titration Experiment with Microarrays

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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.

More Related