GSM7915346: PCNB_T11_K3; Escherichia coli; RNA-Seq
Total RNA was extracted with TRI Reagent (Sigma Aldrich) from 12 samples from the three biologically independent kinetics as follows: for MG1655, T0, 0.5, 2 and 4 min for the first kinetic; T0, 1, 2.5 and 5 min for the second kinetic; and T0, 1.5, 3 and 7 min for the third kinetic; for the pcnB mutant, T0, 1, 3 and 7 min for the first kinetic; T0, 0.5, 1.5 and 5 min for the second kinetic; and T0, 2, 4 and 11 min for the third kinetic. DNA contamination was removed with a Turbo DNase treatment (Invitrogen). Total RNA (1 µg) was ribodepeleted using the riboPOOLs kit (siTOOLs Biotech). 10 ng ribodepleted RNA was further used to construct a sequencing library using the Ion Total RNA-Seq Kit v2 and the Ion Xpres RNA-Seq Barcode Kit (Thermo Fisher Scientific). The library was constructed according to the manufacturer's protocol. After quality control, 15 pg/µl of each library were amplified by emulsion PCR on the Ion One Touch 2 instrument. Then, 100 base pair [bp] single-end library was sequencing on an Ion GeneStudio S5 system with an Ion 540 Chip was performed by the GeT-BioPuces platform (http://get-biopuces.insa-toulouse.fr/).