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Redefining DNA Microarrays

Redefining DNA Microarrays. Tiffany McAninch Advised by: Dr. Frederick Haselton, PhD Mark McQuain, 2BPhD. Purpose. Detect gene expression Understand body and disorders DNA -> mRNA -> protein. Northern Blotting. Microarrays. Same info., but hundreds at a time Whole genetic distribution.

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Redefining DNA Microarrays

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  1. Redefining DNA Microarrays Tiffany McAninch Advised by: Dr. Frederick Haselton, PhD Mark McQuain, 2BPhD

  2. Purpose • Detect gene expression • Understand body and disorders • DNA -> mRNA -> protein

  3. Northern Blotting

  4. Microarrays • Same info., but hundreds at a time • Whole genetic distribution

  5. Microarray Procedure • Target genes prepared and hyb. onto slides • Culture cells • Isolate mRNA • Prepare labeled cDNA • 2 cDNAs competitively hybridized • Analyzed by fluorescent signature

  6. Microarray Schematic

  7. Arrayer • $50,000

  8. Goals • Design system to do same thing • Make quicker • Cheaper • Easier

  9. The Bead Raman Tag DNA Target Microbead cDNA Binding Site

  10. PMT Dichroic Filters PMT Flow Manifold PMT Bandpass Filters SSC Laser FSC Cells Flow Cytometer

  11. Microbeads Easier to store Do chemically Hold patent! Less Reagents Speed Flow Cytometer 2 million signatures Microarrays Don’t need Raman tag No flow cyt. for Raman 50,000 signatures Comparison

  12. Work Completed • Determined right beads and ordered • Learned Raman • Analyzed Raman of test beads and 2 ordered • Got primer for biotinylated targets and did PCR (polymerase chain reaction) • Calculated number of binding sites

  13. Work to do • Hybridize fluorescent target DNA to beads • Hybridize without flur. and meausure Raman • mRNA -> cDNA • Flurescently label and hybridize to target DNA • Attach Raman tags

  14. Thanks!! • Professor Haselton • Mark McQuain • Professor Mahadevan-Jansen • Chad Lieber • Todd Monroe

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