title: GSM5766816: Patient3 breast cancer tissue; Homo sapiens; ncRNA-Seq
library_construction_protocol: Total RNA was extracted and digested using RNase R. Strand-specific RNA-seq libraries were prepared using the VAHTS Total RNAseq (H/M/R) Library Prep Kit for Illumina (Vazyme, Nanjing, China). Briefly, the digested RNA samples was fragmented and then used for first- and second-strand cDNA synthesis with random hexamer primers. In the second-strand synthesis reaction, dTTP was replaced with dUTP. After cDNA synthesis, the double-stranded products were end repaired, a single 'A' base was added, and adaptors were ligated onto the cDNA products. Fragments of cDNA were captured using magnetic particles and treated with uracil DNA glycosylase to remove the second-strand cDNA. Purified first-strand cDNA was subjected to PCR amplification, followed by analysis of the libraries with a Bioanalyzer 4200 (Agilent, CA, USA); the cDNA was then sequenced on an Illumina HiSeq X Ten (X10) system (Illumina, CA, USA).