RNA-seq of Pv11 cells: 24 h of trehalose treatment, rep. 3
Pv11 cells were treated for 24 hours with 0.6M trehalose in water with 10% of complete IPL medium (IPL-41 medium with 10% fetal bovine serum and 2.6 g/l tryptose phosphate broth). 300 g of total RNA isolated with TRIZOL from Pv11 cells were processed using NEB Next Ultra RNA kit. Poly-A mRNA was isolated with 24-dT oligonucleotides, NEB Next Poly(A) mRNA Magnetic Isolation Module, fragmented, reverse transcribed with 6-nucleotide random primers. Second strand cDNA was synthesized, adapters ligated and libraries were amplified using a universal primer (the same for all libraries) and index primers. Then, libraries were cleaned with AMPure XP. Library was quantified with Qubit 3.0, length distribution was estimated with Bioanalyzer. Library was sequenced on Illumina HiSeq 2500 with 50 bp single-end sequencing.