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

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

3. Northern Blotting

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

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

6. Microarray Schematic

7. Arrayer n $50,000

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

9. The Bead

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

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

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

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

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