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

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

Northern Blotting

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

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

Microarray Schematic

Arrayer n $50,000

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

The Bead

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

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

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

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

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