title: GSM7136054: KO clone 2; Mus musculus; RNA-Seq
library_construction_protocol: RNA was extracted from the mpkCCDs using Direct-zol RNA MicroPrep kit. Trizol-extracted RNA was eluted in 35 µl DNA/RNA free water. First-strand cDNA was prepared from 50 ng of total RNA using SMART-Seq v4 Ultra Low Input RNA Kit, followed by cDNA amplification with 9 PCR cycles. 1 ng of the synthesized cDNA was fragmented and tagged with index primers using the Nextera XT DNA Sample Preparation Kit (Illumina) to generate sequencing libraries for high throughput sequencing. After the purification of libraries with AmPure XP magnetic beads (Beckman Coulter), each library was quantified using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific). An equal amount of the index libraries were pooled, and the pooled libraries were sequenced on an Illumina platform (50-bp paired-end NovaSeq).