RNA-seq of Pv11 cells: control, rep. 2
Pv11 cells cultivated in accordance with standard protocol: IPL-41 medium with 10% fetal bovine serum and 2.6 g/l tryptose phosphate broth, incubation in non-humidified incubator at 25 C. 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.