title: GSM8648510: H9-DDdCas9VP192-FOXL2 clone 24 cells, day 8, induced, replicate 3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using the Nucleospin RNA kit (Macherey-Nagel) skipping the DNAse treatment steps. Seperated DNAse treatment was performed with the RQ1 RNAse-free DNAse (Promega) to remove gDNA after which the RNA was purified using the Nucleaspin RNA Clean-up kit (Macherey-Nagel). RNA libraries were prepared by Novogene. Messenger RNA (mRNA) was isolated from tRNA employing poly-T oligo-attached magnetic beads (ABclonal) for mRNA library preparation. The mRNA was then fragmented and thereafter cDNA was synthetised through reverse transcription. The first cDNA strand was synthetised using random hexamer primers and the second cDNA strand with dUTP/dTTP. The non-directional library was ready after end-repair A-tailing, adapter ligation, size selection, amplification and purification. The mRNA library was subsequently quantified with Qubit (Invitrogen) and RT-PCR. Bioanalyzer was used to check the size distribution detection.