GSM1876371: MabATCC_tss_R2; Mycobacteroides abscessus ATCC 19977; RNA-Seq
Following physical disruption using 0.1 mm zirconia-silica beads in a MagNA Lyzer (Roche Diagnostics), RNA was isolated as previously described (doi: 10.1111/mmi.12387). Briefly, RNA was isolated using miRNeasy spin columns with an additional on-column DNase treatment, according to manufacturer's instructions (Qiagen). Two types of strand specific libraries were prepared in parallel, mRNA enriched (TEX+) and not enriched (TEX-) libraries, and sequenced using the Illumina HiSeq 2000 instrument in a single read mode. Briefly, for mRNA enriched samples, monophosphorylated i.e. processed transcripts were removed from total RNA by a 1 h treatment at 30 °C with a terminator-5′-phosphate-dependent exonuclease (TEX+) (TEX, Epicentre Biotechnologies). The not enriched samples (TEX-) were incubated in the reaction buffer alone, without the enzyme. Obtained RNA samples were then treated with a tobacco acid pyrophosphatase (TAP, Epicentre Biotechnologies) at 37°C for 1 hour, followed by a 5' linker ligation and synthesis of cDNA.
GEO Accession
GSM1876371