Library construction and sequencing
Isolated RNA was separated into two fractions. The first fraction
included eukaryotic mRNA that was extracted from total RNA by NEBNext®
Poly(A) mRNA Magnetic Isolation Module (BioLabs) following
manufacturers’ protocol. The second fraction (bacterial RNA and
eukaryotic rRNA) was extracted from the supernatant after mRNA
separation by RNAClean XP beads (Beckman Coulter). Both fractions were
used for library constructions. cDNA libraries were built by Zymo-Seq
RiboFree Total RNA-Seq Library Kit (Zymo Research) following
manufacturers’ protocol. The quality of cDNA libraries was then
visualized on Bioanalyzer by High Sensitivity DNA Kit (Agilent).
Libraries were sequenced at CEITEC institute (Brno, Czechia) on the
Illumina novaseq platform on a 2x 150 bp paired-end reads run.