title: GSM2054452: wild type strain, H37Rv, treated with 3.9 μM of compound 14; Mycobacterium tuberculosis; RNA-Seq
library_construction_protocol: Equal volume of 4M guaidinium thicyanate was added to the bacterial suspension and the pellet collected by centrifugation. The pellet was suspended in 1mL of Trizol and transferred to scewcap tubes containing ~0.5 mL of zirconia beads. The cells were lysed by bead-beating (3 cycles) and the lysate collected by centrifugation. Chloroform extraction was performed on the lysate and the aqueous fraction transferred to RNAeasy columns (Qiagen) for purification of RNA. The RNA sample was subjected to DNase (Ambion) digestion for 1h and then treated with the MicrobExpress bacterial mRNA purification kit (Ambion) to remove contaminating rRNA and tRNA. The enriched samples were then used for library construction. TruSeq RNA sample preparation kit v2 from Illumina was used to generate the libraries by following the Low Sample (LS) protocol. The following adaptors were used: 1) Sample 1: AR005 2) Sample 2: AR004 3) Sample 3: AR012 4) Sample 4: AR019 5) Sample 5: AR002 6) Sample 6: AR004.
attributes.tag: GEO Accession
attributes.value: GSM2054452