title: GSM6514076: Prostate cancer tissue, T1-T2, 1 [S1-1]; Homo sapiens; ncRNA-Seq
library_construction_protocol: Total RNA was extracted from the test samples by Trizol method, and the samples were firstly analyzed for DNA contamination and integrity by agarose gel electrophoresis; then the RNA concentration and purity (OD260/280) were detected by Nanodrop, and if the OD260/280 values were in the range of 1.8-2.1, the RNA purity was qualified; next, the RNA purity was detected by Agilent 2100. Agilent 2100 was used to detect the purity and integrity of the total RNA. The sequencing of the test RNA samples was entrusted to Shanghai Jikai Gene Technology Co 5ug of each sample was prepared for library preparation. Ribosomal rRNA was removed using the Ribo-zero™ rRNA Removal Kit (Epicentre, USA), then eliminated linear RNA using RNase Rase. The remaining circRNA was broken into short fragments of 250-300 bp, which were used as a template to synthesize the second strand of cDNA using random oligonucleotides as primers to synthesize the first strand of cDNA, followed by dNTPs (dUTP, dATP, dGTP, dCTP) as raw materials to synthesize the second strand of cDNA. The purified double-stranded cDNAs were end-repaired, A-tailed, and sequenced, and AMPure XP beads were used to screen 350-400 bp cDNAs in length. cDNAs containing U were degraded with USER enzymes, and PCR amplified to obtain libraries. The fragment size and purity of the library were checked using Agilent 2100 to ensure the quality of the library. Illumina PE150 double-end sequencing was performed after passing quality control.