GSM8329271: CMP2; Mus musculus; ncRNA-Seq
Total RNA was isolated using Trizol (Invitrogen). 1~3 μg total RNA per sample was used for library construction, which was generated using NEBNext Multiplex Small RNA Library Prep Set for Illumina (NEB) following manufacturer's instructions. After synthesis of first strand cDNA, PCR amplification was performed using LongAmp Taq 2X Master Mix, SR Primer for illumina and index primer. PCR products were purified on a 8% polyacrylamide gel (100V, 80 min). DNA fragments corresponding to 170~370 bp (the length of snoRNA plus adaptors) were recovered and dissolved in 8 μL elution buffer. At last, library quality was assessed on the Agilent Bioanalyzer 2100 system using DNA High Sensitivity Chips. After clustering of the index-coded samples, the library preparations were sequenced on an Illumina NovaSeq platform.