GSM8499131: Synovial fibroblast, siDLX4a; Homo sapiens; RNA-Seq
Total RNAextracted by Qiagen RNAeasy kit, and quantified by Nanodrop. 400 ng per sample was sent to Novogene (Beijing, China) for sequencing and analyses Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using randomhexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional libraryor dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, sizeselection, amplification, and purificationFor the directional library, it was ready after end repair, A-tailing, adapter ligation, sizeselection, USER enzyme digestion, amplification, and purification.The library was checked with Qubit and real-time PCR for quantification and bioanalyzer forsize distribution detection. Quantified libraries will be pooled and sequenced on Illuminaplatforms, according to effective library concentration and data amount. The clustering of the index-coded samples was performed according to the manufacturer'sinstructions. After cluster generation, the library preparations were sequenced on an Illuminaplatform and paired-end reads were generated