Cellular active small molecule inhibitors of Mycobacterium tuberculosis by NMR fragment screen

Cellular active small molecule inhibitors of Mycobacterium tuberculosis by NMR fragment screen. Microarray experiments were performed as dual-color hybridizations on reporter level. In order to compensate specific effects of the dyes and to ensure statistically relevant data analysis, a color-swap dye-reversal was performed. Two different labeling methods were applied. RNA labeling was performed with the two color Quick Amp Labeling Kit (Agilent Technologies) using FullSpectrum MultiStart Primer for T7 IVT RNA Amplification (BioCat GmbH, Heidelberg, Germany) as random T7 labeling. Alternatively, the total RNA samples were amplified with the TransPlex Whole Transcriptome Amplification Kit (Sigma-Aldrich, Munich, Germany) and labeled with BioPrime Plus Array CGH Indirect Genomic Labeling System (Invitrogen, Karlsruhe, Germany) as WTA BioPrime indirect. Both methods were combined in the data analysis and only results derived from both labeling methods were considered as relevant for further verification.