GSM5832828: N1-circ; Homo sapiens; RNA-Seq
The peripheral blood samples and TRizol reagent(Invitrogen, USA) are evenly mixed in a ratio of 1:3, total of 12 ml. Total RNA was extracted by using Trizol reagent according to the manufacturer's instructions. Whole transcriptome libraries were constructed by the TruSeq Ribo Profile Library Prep Kit (Illumina, United States), according to the manufacturer's instructions. In brief, 10 mg total RNA was depleted rRNA with an Illumina Ribo-Zero Gold kit and purified for end repair and 50-adaptor ligation. Then, reverse transcription was performed with random primers containing 30 adaptor sequences and randomized hexamers. Finally, the cDNAs were purified and amplified with thermo cycler. The PCR products of 300-500 bp were purified, quantified and stored at -80℃ before sequencing. The libraries were subjected to 151 nt paired-end sequencing with an Illumina Nextseq 500 system (Novogene, China). For small RNA (sRNA) sequencing, eight sRNA libraries were generated with TruSeq small RNA (Illumina, United States) according to the manufacturer's instructions. Then the prepared libraries were sequenced with an Illumina Novaseq 6000 system (Novogene, China).