GSM5729303: ko-3; Mus musculus; RNA-Seq
Two micrograms of total RNA were used for RNA sample preparation. Sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer's recommendations and index codes were added to attribute sequences to each sample. RNA concentration of library was measured using Qubit® RNA Assay Kit in Qubit® 3.0 to preliminary quantify and then dilute to 1 ng/μl. Library quality was assessed using the Agilent Bioanalyzer 2100 system. cDNA was preamplification for 16-18 cycles by using KAPA HiFi HotStart ReadyMix (KK2601, Roche) and IS PCR primers, followed by PCR purification by AMPure XP beads (A63881, Beckman). Finally, amplified cDNA was fragmented and RNA-seq library was constructed by using TruePrep DNA Library Prep Kit (TD502, Vazyme) according to the manufacturer's instructions. Paired-end 150-bp sequencing was performed on the Illumina HiSeq X-Ten or NovaSeq 6000 system.