Thursday, June 28, 2012

Improving Microarray Hybridization

Arrayit Array Plate Multi-Well Hybridization Station accelerates hybridization rates by approximately 10-fold compared to static cover slip hybridization reactions. Two genotyping microarrays were hybridized for 30 minutes at 42° in 30 µl under a glass cover slip (top) or in 200 µl at 500 rpm using an Array Plate Hybridization Station (bottom). Two-color red-green probes mixtures were prepared in 1X UniHyb® 2 Hybridization Solution at final probe concentration of 1.6 nM (top) and 0.2 nM (bottom). The microarrays were washed for 5 min at 45° in Wash Buffer 1, 5 min at room temperature in Wash Buffer 2, 1 sec in Wash Buffer 3, spun dry for 3 sec in a Microarray High-Speed Centrifuge and scanned using an Arrayit InnoScan® 710AL Two-Color High-Speed Fluorescence Scanner at high laser power and gain 10 in the red channel and gain 5 in the greeen channel. The Array Plate produced stronger signals even with the 6.7-fold greater probe dilution, indicating an approximately 10-fold increase in hybridization signal with 500 rpm Array Plate mixing.