Primers (Invitrogen) were designed for all 1738 open reading frames (ORFs) using Primer 3 software. Polymerase chain reaction was performed with 50 ng of genomic DNA, 0.8 uM of each forward and reverse primer, and 10 U of Taq DNA polymerase (New England Biolabs). Products were precipitated on 96 well plates with isopropanol and 3M sodium acetate at 4C and analyzed on a 0.8% agarose gel. Poly-L-lysine SuperChip slides (Erie Scientific) were printed using a homemade linear servo arrayer. For more details, please see the documentation provided by the Derisi Lab on their homepage at UCSF. Dried PCR products were resuspended in 3X SSC and rearrayed in 384 well plates. Three copies of each ORF were printed in two sub-arrays on the slides for a density of 6018 PCR spots (total of 7040 spots) on each sub-array.
Support
glass
Coating
polysine
Description
Each sub-array contains blocks arranged 4 by 4 with each block running 22 rows by 20 columns.
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