5'-RNA-seq of 5'-enriched primary coding RNA in Staphylococcus aureus wildtype to detect Transcription Start Sites

ABSTRACT: To detect Transcription Start sites (TSS) in Staphylococcus aureus wildtype a cDNA library enriched for primary 5′-transcripts according to Peifer-Sancar et al. (2013) was prepared and sequenced on Illumina MiSeq system to retrieve 5′-ends of primary transcripts. Cultivation was performed in Luria Bertani (LB) medium in triplicate at 37°C until cells have reached an optical density at 540 nm of 2.0. Cells were harvested by centrifugation, washed with Belitsky minimal medium (BMM) and adapted to BMM for one hour. S. aureus cells of 3 replicate experiments were harvested and disrupted in 3 mM EDTA/ 200 mM NaCl lysis buffer with a Precellys24 Ribolyzer. RNA isolation was performed using the phenol-chloroform-isoamylalcohol approach. After precipitation with 3 M sodium acetate and isopropanol

INSTRUMENT(S): Illumina MiSeq

ORGANISM(S): Staphylococcus aureus subsp. aureus

SUBMITTER: Tobias Busche 

PROVIDER: E-MTAB-7385 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress