GSM9753172: NC1; Mus musculus; RNA-Seq
Total RNA was extracted using the TransZol Up reagent (TransGen Biotech) according to themanufacturer's instructions. After extraction, the RNA integrity was checked using 1.5% agarose gel electrophoresis or a fragment analyzer. The concentration and purity of the RNAwere measured using a Nanodrop spectrophotometer. The transcriptome library was constructed using the VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina according to the manufacturer's protocol. The steps are as follows: 1. Add fragmentation buffer and random primers to fragment the target RNA and allow random primers to bind to the target RNA. 2. Add first-strand reverse transcription reagents to reverse transcribe the target RNA into cDNA, forming RNA-cDNA hybrids. 3. Add second-strand synthesis reagents to cleave the RNA from the RNA-cDNA hybrids and use the remaining RNA as primers to synthesize double-stranded cDNA. 4. Perform end-repair and add an A-tail to the double-stranded cDNA, then ligate Y-shaped adapters. 5. Use magnetic beads to remove adapter dimers. 6. Enrich the library templates using PCR amplification. 7. Recover the library using magnetic beads.