GSM6603322: Old_Extratumoral tissue_Naive; Mus musculus; RNA-Seq
Total RNA was isolated with Trizol reagent and a PureLink RNA Mini Kit (Ambion 12183025). Messenger RNA was purified from total RNA using poly-Toligo-attached magneticbeads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, sizeselection, USER enzyme digestion, amplification, andpurification. The clustering of the index-coded samples was performed according to the manufacturer's instructions. After cluster generation, the library preparations were sequenced on an Illumina platform and paired-end reads were generated.