We produced in-house DNA microarrays that contain PCR-amplified fragment of DNA derived from predicted open reading frames (ORFs) in the E. coli genome. All PCR products were analyzed by electrophoresis on 1% agarose gels. Reactions that resulted in undetectable PCR products were re-amplified using a lower annealing temperature. All PCR products from the final round of amplifications were precipitated with ethanol, then dissolved in 50% DMSO (100-150 μg/ml) and spotted onto Corning GAPS II slides using Affymetrix GMS 417 Arrayer (Santa Clara, CA) in duplicate on each slide. Arrayed slides were baked at 80˚ C for 2 hours and stored in a desiccator at room temperature under vacuum.
Description
The Platform data table reflects a condensed representation of the duplicate-spotted PCR products on the physical array.
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