GSM5135412: PSD1: Planktonic-shedding_Rep1; Burkholderia pseudomallei K96243; RNA-Seq
Total RNA was extracted from bacterial pellets and performed using a TRIzol reagent with the Purelink RNA mini kit (Ambion, USA). RNA integrity was evaluated using Agilent 2100 Bioanalyzer (Agilent Technologies, USA). The total RNA sample 2.5 μg was used to construct the RNA libraries. Total RNA in each sample, after removal of the 23S and 16S ribosomal RNA (rRNAs) with Ribo-Zero Gram-negative Bacteria rRNA removal kit (Epicentre Technologies, USA), was purified by an ethanol precipitation. The cDNA libraries were assembled from the fragmented purified rRNA-depleted RNA using a ScriptSeq v2 RNA-Seq Library Preparation kit (Epicentre Technologies, USA) and purified using a QIAquick PCR purification kit (QIAGEN, USA). An Illumina Indexed-adapter sequence was ligated to di-tagged cDNA to generate the amplified libraries as per the RNA sample preparation protocol (Illumina, USA). Excess nucleotides and PCR primers were eliminated from the libraries with a QIAquick PCR purification kit and the quantity and quality were examined using an Agilent 2100 Bioanalyzer. cDNA libraries were prepared and constructed using a ScriptSeq v2 RNA-Seq Library Preparation kit (Epicentre Technologies, USA)