GSM2536245: Stationary_Phase_RNA-seq; Caulobacter vibrioides; RNA-Seq
2ml of Caulobacter was harvested in the presence of 1ml 95% ethanol/5% phenol and 8ml PYE. The cell pellets were extracted by hot phenol/chloroform method. Libraries were constructed as detailed previously (Guo et al., 2014). Briefly, ribosomal RNA removed with the MICROBExpress kit (Life Technologies). Samples were lightly fragmented with RNA fragmentation reagents (Ambion), isolating and dephosphorylating 25-50 mers with T4 PNK (New England Biolabs), ligating linker-1 (Integrated DNA Technologies) to fragments with T4 RNA ligase II (Epicentre). After reverse transcription was performed with Superscript III (Invitrogen) and RT_oligo, the RNA was hydrolyzed with NaOH, and the ssDNA cDNA was circularized with CircLigase (Epicentre). Final sequencing libraries were PCR amplified with Phusion polymerase and adapter oligos (o231 and o230).