title: Sample 1
organism: Mycobacterium tuberculosis H37Rv
description: Protocols: Mycobacterium tuberculosis H37Rv (SysteMTb) was grown in Middlebrook 7H9 medium supplemented with 0.4% glycerol, 0.085% NaCl, 0.5% BSA and 0.05% Tyloxapol in roller bottle culture (2 rpm at 37°C). RNA was isolated from exponentially growing cultures using the FastRNA Pro blue kit from QBiogene/MP Bio according to manufactures instructions. RNA samples was treated with Turbo DNase free (Ambion). RNA was fragmented with ultrasound (4 pulses of 30 s at 4oC), then treated with Antarctic phosphatase and re-phosphorylated with polynucleotide kinase (PNK). Fragmented RNA was then poly(A)-tailed using poly(A) polymerase and a RNA adapter was ligated to the 5'-phosphate of the RNA. First-strand cDNA synthesis was performed using an oligo(dT)-adapter primer and M-MLV reverse transcriptase. The resulting cDNA was PCR-amplified to about 20-30 ng/µl using a high fidelity DNA polymerase.
attributes.tag: ENA first public
attributes.value: 2015-01-23
attributes.tag: ENA last update
attributes.value: 2015-01-20
attributes.tag: External Id
attributes.value: SAMEA3214866
attributes.tag: INSDC center alias
attributes.value: National Institute for Medical Research
attributes.tag: INSDC center name
attributes.value: National Institute for Medical Research
attributes.tag: INSDC first public
attributes.value: 2015-01-23T17:45:21Z
attributes.tag: INSDC last update
attributes.value: 2015-01-20T21:23:47Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-3252_2:Sample 1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: sample name
attributes.value: E-MTAB-3252_2:Sample 1