title: GSM6297512: grp4_rep2; Rattus norvegicus; RNA-Seq
library_construction_protocol: From frozen pellets of the cultured cells (>10**6), total RNA was extracted with NucleoSpin RNA (Macherey-Nagel GmbH & Co. KG, Düren, Germany). Using the total RNA as a template (1.0 ng), the double stranded cDNA was synthe-sized by SMART (Switching Mechanism At 5'End of RNA Template) method with SMART-Seq® v4 Ultra® Low Input RNA Kit for Sequencing (Clontech Laboratories, Inc., CA, U.S.A.). The double stranded cDNA was amplified in 13 cycles of PCR. The PCR product was purified by the magnetic bead method with AM Pure XP (Beckman Coulter, Inc., CA, U.S.A.). With Nextera XT DNA Library Prep Kit and Nextera XT Index Kit v2 (Illumina, Inc., CA, U.S.A.), the double stranded cDNA was fragmented by tagmentation reaction using transposon and then both ends of the fragmented cDNA was ligated with adapter sequences. The ligated cDNA (0.2 ng) was amplified in 12 cycles of PCR using the primers having a unique sequence for each sample. The PCR products for all samples were pooled into one library.