GSM3666820: RNA sequencing parental cells Rep 1; Mycobacterium tuberculosis H37Rv; RNA-Seq
The bacterial pellets were resuspended in Trizol and RNA was isolated and purified using RNAeasy columns. The purified RNA was shipped to Aggrigenome Labs Pvt Ltd (India). RNA integrity was checked using Agilent RNA 6000 Nano Kit (Agilent Technologies, Santa Clara, CA, USA). Ribosomal RNA was removed from total RNA sample using Ribo-ZeroTM Magantic kit (Bacteria) as per manufacturer's recommendations (Epicentre). RNA cleanup was performed using Agencourt RNACleanTM XP Beads (Beckman Coulter). RNA was fragmented followed by random priming for first strand cDNA synthesis and second strand synthesis. Adapters were ligated and the library was cleaned up twice using Agencourt RNACleanTM XP Beads followed by 14 PCR amplification cycles. Amplified library was further cleaned up using AMPure XP Beads (Beckman Coulter Inc). The quality of library was checked using Standard Sensitivity NGS Fragment Analysis Kit on an Advanced Analytical Fragment Analyzer. Library was quantified using qPCR as per Illumina Sequencing Library qPCR quantification guide and sequencing was performed at Illumina HiSeq2000.
GEO
GSM3666820