Tribromsalan 2
Mycobacterium tuberculosis H37Rv
Protocols: Mycobacterium tuberculosis H37Rv was cultured in Middlebrook 7H9 broth (with 0.05% Tween 80 and 10% albumin-dextrose-catalase). Drugs were prepared as 10 mM stock solutions in DMSO and stored at -20'C in aliquots to avoid freeze-thawing. Log phase M. tuberculosis cultures (OD 0.2-0.4, 4-day shaking at 150 rpm) were treated with 10x MIC niclosamide (final concentration 30.9 uM) or tribromsalan (final concentration 681.6 uM) for 4 hours, or vehicle control only (sterile DMSO, final concentration 0.75%), from three independent biological replicates on different days. Following drug exposure, mycobacterial RNA was extracted using an established GTC/TRIzol method (Waddell et al., Tuberculosis 2004; 84:263-274). The resulting mycobacterial RNA was DNase-treated and purified using the RNeasy spin columns (Qiagen, Hilden, Germany). RNA yield and quality were assessed by NanoDrop One (Thermo Scientific, Waltham, MA, USA) and Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). RNA yields from all samples were similar and of good quality. Mycobacterial ribosomal RNA was depleted using RiboCop META kit (Lexogen, Vienna, Austria) resulting in mRNA suitable for RNA sequencing. Strand-specific RNAseq libraries were prepared using the NEBNext Ultra II directional RNA library prep kit (New England Biolabs, Ipswitch, MA, USA) and pooled in-house.
ENA first public
2024-03-01
External Id
SAMEA115172679
INSDC center name
Department of Global Health and Infection
INSDC last update
2024-02-02T09:43:51Z
INSDC status
public
Submitter Id
E-MTAB-13761:Tribromsalan 2
broker name
ArrayExpress
collection date
not collected
geographic location (country and/or sea)
not collected
growth condition
Log phase in vitro
isolate
not applicable
organism part
whole organism
replicate
Biological Replicate 2
sample name
E-MTAB-13761:Tribromsalan 2
scientific_name
Mycobacterium tuberculosis
strain
H37Rv
type-material
type strain of Mycobacterium tuberculosis