title: RNA-Seq of wild type: Biological Repeat 2
design: The purified RNA was used for the preparation of the sequencing library using TruSeq Stranded mRNA Library Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturers instructions. Briefly, mRNA was purified from total RNA (1 m) by oligo(dT)-coupled magnetic beads and fragmented into small pieces under elevated temperature. The first-strand cDNA was synthesized using reverse transcriptase and random primers. After generating double-strand cDNA and adenylation on 3 ends of DNA fragments, adaptors were ligated to the ends of the fragments. The products were enriched with PCR and purified with AMPure XP system (Beckman Coulter, Beverly, USA). The libraries were analyzed and qualified by a Qsep400 Bio-Fragment Analyzer (Bioptic Inc., New Taipei City, Taiwan) and a Qubit 2.0 Fluorometer (Thermo Scientific, Waltham, MA, USA). The qualified libraries were then sequenced on an Illumina NovaSeq platform with 150 bp paired-end reads generated by Genomics BioSci & Tech Co., New Taipei City, Taiwan.