GSM1647529: C. lari 46°C Replicate 1; Campylobacter lari; RNA-Seq
Total RNA was extracted using a hot phenol/lysozyme method The libraries were generated by Vertis Biotechnologie AG (Munich, Germany). The RNA samples were poly(A)-tailed by using poly(A) polymerase. The 5'PPP were removed using tobacco acid pyrophosphatase (TAP) followed by the ligation of the RNA adapter to the 5'-monophosphate of the RNA. First-strand cDNA synthesis was performed with an oligo(dT)-adapter primer and the M-MLV reverse transcriptase. The resulting cDNA was PCR- amplified to reach a concentration of 20-30 ng/μl using a high fidelity DNA polymerase. The cDNA was purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics, Essex, UK) and was analyzed by capillary electrophoresis. The primers used for PCR amplification were designed for TruSeq sequencing according to the instructions of Illumina (San Diego, CA).