GSM9141490: Old-OSK-rep3; Mus musculus; RNA-Seq
Total RNA was extracted following the TRIzol Reagent User Guide (Thermo Fisher Scientific). Glycogen (1 μl of 10 mg ml−1 solution) was added to the supernatant to increase RNA recovery. RNA was quantified using a Qubit 2.0 Fluorometer (Life Technologies), and RNA integrity was determined using TapeStation (Agilent Technologies). RNA library preparations, sequencing reactions were conducted at Genewiz. A SMART-Seq v4 Ultra Low Input Kit for Sequencing was used for full-length cDNA synthesis and amplification (Clontech), and Illumina Nextera XT library was used for sequencing library preparation. In brief, cDNA was fragmented and adaptors were added using Transposase, followed by limited-cycle PCR to enrich and add an index to the cDNA fragments. The final library was assessed by a Qubit 2.0 Fluorometer and an Agilent TapeStation.