GSM1692726: Oxidative stress 90min BW25113; Escherichia coli BW25113; ncRNA-Seq
Total RNA was extracted using Trizol method. tRNAs were deacylated at pH=10. Poly(A) tail was added to the deacylated 3’-CCA terminus by E. coli poly(A) polymerase (NEB) according to the manual. Prior to reverse transcription, polyadenylated tRNA were denatured with 100 µM TG primer (5’-T15GG-3’) and dNTP (25mM each) at 80°C for 5 min and then chilled on ice. The reverse transcription was performed with RevertAid Premium Reverse Transcriptase (Fermentas) according to the manual. After transcription of tRNA to cDNA, the second-strand synthesis was performed with RNase H, DNA Polymerase I and T4 DNA Polymearse (Fermentas) according to the manuals. The sequencing library was constructed using NEBNext Ultra DNA library prep master mix set for Illumina (NEB). Pair-end (sequenced in single-end)
GEO Accession
GSM1692726