title: GSM2526490: RNA_cid16Δ; Schizosaccharomyces pombe; RNA-Seq
library_construction_protocol: Total RNA was isolated applying the hot phenol method. The extracted nucleic acids were treated with DNAse I for 2 h at 37°C. DNAse was removed by a second phenol-chloroform-isoamylalcohol extraction and ethanol precipitation. 1 µg total RNA was degraded with Terminator nuclease (Epicentre) in buffer A at 30°C for 2 h. RNA libraries were prepared with NEBnext Ultra Directional RNA Library Prep Kit for Illumina (NEB); DNA library construction with NEBNext Ultra II DNA Library Prep Kit for Illumina kit (NEB). siRNA library: ligation of 3' adapter (5'-App CTG TAG GCA CCA TCA AT/ddC/-3') and 5' adapter (5'-GUU CAG AGU UCU ACA GUC CGA CGA UC-3'), reverse transcription, PCR of cDNA with Illumina P5 5' primer and barcoded llumina P7 3' primer.