GSM2732741: HC-1; Homo sapiens; RNA-Seq
Total RNA was extracted from each sample of PBMCs by Trizol reagent (Invitrogen, USA) according to the protocol of manufacturer, and treated with RNase-free DNase to remove traces of genomic DNA. For high-throughput sequencing, we synthesized the cDNA libraries of the circRNA from each sample based on the Illumina standard protocols. Briefly, total RNA was treated with the Epicentre Ribo-Zero kit (Epicentre, USA) to remove all the ribosomal RNAs, and digested with RNase R (Epicentre, USA) to remove all the linear RNAs, thus got the pure circular RNAs. Then, the enriched circular RNAs were processed by the NEBNext® Ultra™ Directional RNA Library preparation kit (Illumina, USA) according to the Illumina protocol. The main steps involved: the fragmentation of circRNA, reverse transcription and second-strand cDNA synthesis, end repair, dA-tailing and adaptor ligation. The products of these reactions were purified and amplified by PCR to create the final circular RNA cDNA library. The libraries were paired-end sequenced on the Illumina HiSeq2500 platform (Illumina, USA).
GEO Accession
GSM2732741