GSM9009610: Control, mouse dermal fibroblasts -3; Mus musculus; RNA-Seq
RNA was extracted from primary dermal fibroblasts using RNeasy Plus Micro Kit (Qiagen, cat. no. 74034) according to manufacturer's protocol. Paired-end sequencing was performed at Novogene (UK) on Illumina platform (PE150, 20 million reads). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using dTTP for non-directional library. The non-directional library was complete after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.