GSM5665207: Sirt55; Mus musculus; RNA-Seq
Ribosomal RNA (rRNA) was removed from Total RNA using QIAseq FastSelect RNA Removal Kit (Qiagen, Venlo, The Netherlands) according to the manufacturer's instructions. The enriched mRNAs and non-coding RNAs were fragmented into short fragments (approximately 300 bp) using a fragmentation buffer and reverse-transcribed to cDNA with random primers, and this first-strand cDNA was used as the template for second-strand cDNA synthesis. Then, the cDNA fragments were end-repaired and ligated to Illumina sequencing adapters. The second-strand cDNA was digested using Uracil-DNA Glycosylase (UDG). The digested products were size selected (approximately 450 bp), PCR amplified, and quality assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA). After cDNA library construction, a paired-end sequencing was performed using Illumina Hiseq 4000 systems (Illumina, San Diego, CA, USA). The raw data were filtered, and the clean data were mapped to the reference genome Mus_musculus. GRCm38. RNA libraries were prepared for sequencing using standard Illumina protocols