A-MAXD-2 - EhV86 MICROARRAY
EhV86 microarray Version 1 is desgned in a 4 X 4 metagrid, with each subgrid containing features printed in 12 rows of 13 columns. Microarray slides (Corning GAPS II Amino-propyl-silane) were printed using BioRobotics MG2 Model 14127. Pins used were 16x BioRobotics Microspot 2.5K. 50uM of each oligo was spotted (a 1/6 dilution of 300uM stocks) in 150mM phosphate buffer including 0.01% SDS.
Spotting Method: Post-processing and Prehybridisation Slides rehydrated by placing printed side down in a humidity chamber (1X SSC) for 5 mins, then transferred to a hotplate (70oC) while preparing NMP solution. After vigourous dunking slides were left for 15 mins in the freshly made NMP solution (3g succinic anhydride, 175ml NMP, 7.5ml 1M borate buffer) . Slides were dunked 5 times and then left for 2 mins in boiling water, dunked 5 times in absolute ethanol and then centrifuged (1000g, 1 min). Dry slides were then UV cross linked (3000J/s) and transferred into 200ml preheated BSA blocking solution (160ml water, 7.6ml 30% BSA stock, 55ml 20X SSC, 2.2ml 10% SDS). Slides were incubated in BSA blocking solution at 42oC for 1 hour with gentle shaking. Following the prehrybridisation incubation, slides were dunked 5 times in water, then 5 times in isopropanol, centrifuged (1000g, 1 min) and used immmediately.
Adhesion Method: UV crosslinking
|All experiments done using A-MAXD-2: (E-MAXD-11, E-MAXD-2, E-MAXD-23, E-MAXD-8)|