GSM3175263: 5A18-NP1 rep2; Borreliella burgdorferi; RNA-Seq
Three independent cultures of each of WT and MTase mutant strains were grown to mid-late log phase (3 X 107 cells / ml) for RNA isolation. RNA was isolated by first adding 20 ml RNAprotect (Qiagen) to 10 ml culture to stabilize transcripts during processing. Cells were collected by centrifugation, resuspended in 1 ml of pre-warmed (65°C) Trizol reagent (Invitrogen), and frozen at -80°C overnight. Cell suspensions were thawed at room temperature, and RNA was isolated using the Direct-zol RNA mini kit (Zymo Research) following the manufacturer's instructions. RNA concentration was determined using using a Qubit® 2.0 Fluorometer (Life Technologies), and RNA integrity was verified using microfluidic-based capillary electrophoresis with an Agilent 2100 Bioanalyzer (RIN ≥ 9.8 for all samples). Directional cDNA libraries were prepared from 5 μg of purified RNA using the ScriptSeq complete bacteria kit and SciptSeq index PCR primers (Illumina)  following the manufacturer's instructions.
GEO Accession
GSM3175263