title: RNA-Seq analysis of cell suspension culture of Hypericum perforatum
design: RNA-Seq analysis of cell suspension culture of Hypericum perforatum
library_name: Sample 6_p
library_construction_protocol: After co-cultivation of Agrobacteria and A. tumefaciens, the cells were collected at different time points (0.5 h, 3 h, 12 h, 24 h). After the cells were vacuum filtered from the medium, they were immediately cryopreserved in liquid nitrogen and stored at -80 °C.  Three biological replicates for the treatment and control were incubated in a growth chamber with a photoperiod of 16 hours of light and 8 hours of darkness at 25 °C, an irradiation of 80 µmol m-²s-¹ and a relative humidity of 70, and both the treated and control cultures were kept alive. A. tumefaciens or A. rhizogenes was added to H. perforatum cell suspension cultures in the exponential phase (three to five days after subculture).  Total RNA was isolated from 100 mg of biomass using the Sigma Spectrum RNA extraction kit (Sigma-Aldrich, USA). RNA samples with an RNA integrity number (RIN) of at least seven were used for RNAseq analysis.  Random hexamer primers were used to synthesise the cDNA after random fragmentation of the mRNA. DNA polymerase I, dNTPs, RNase H and a special second-strand synthesis buffer (Illumina) were used to generate the second strand. 
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-15161:Sample 6_p