bugs.sgul.ac.uk:BuG@Sbase/Protocol:1195 - bugs.sgul.ac.uk:BuG@Sbase/Protocol:1195
Each hybridization mixture was made up from 500 ng test (Cy3) and 500ng common reference (Cy5) material. Hybridization mixtures were prepared as described in the Agilent Two-Color Microarray-Based Gene Expression Analysis guide version 5.5 (G4140-90050, Agilent technologies) without the inclusion of the RNA fragmentation mixture. M. tuberculosis whole genome arrays were supplied by the Bacterial Microarray Group at St. Georges Hospital, University of London. Hybridization samples were loaded onto 8x15k M. tuberculosis microarrays (Design ID: 027543, Agilent Technologies) and hybridized for 17 hours at 65C. Afterwards the slides were washed and scanned (20 bit, 5um resolution) in an ozone-free room with the Agilent G2505C scanner as described in the Agilent G4140-90050 guide. Data were extracted with Feature Extraction (v10.7.3.1, Agilent Technologies) with the GE2_107_Sep09 protocol for two-color Agilent microarrays.