GSM8457533: NC2 circRNA (biol rep2); Homo sapiens; ncRNA-Seq
Total RNA was isolated from cells using TRIzol (Takara). Total RNA was quantified and qualified by Agilent 2100/2200 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA).1.1 ug of total RNA was used for the construction of sequencing libraries. Total RNA were digested with RNase R to remove linear RNA, followed by Ribomoval rRNA Depletion Kit (Human/Mouse/Rat) (Geneseed,Guangzhou). The designed DNA probe was hybridized to the RNA sample, thereby removing rRNA from the total RNA. Then, the RNA was fragmented and the first strand cDNA was synthesized using Rna-Seq Library Prep Kit for Illumina (Geneseed,Guangzhou). The second strand cDNA was labeled with dUTP in a strand-specific method. At the same time, the end repair is completed in this step. Then, the circRNA-Seq library was obtained by adding A-tail, connecting adaptor, purification of ligation product, fragment size sorting, and library amplification.