GSM3610222: h_MeCP2_KO_H4_102; Homo sapiens; RNA-Seq
Total RNA was isolated from all generated cell lines at day 9 of differentiation using either an RNeasy Mini kit or an AllPrep DNA/RNA Mini kit (Qiagen). Genomic DNA contamination was removed with a DNA-free kit (Ambion) and remaining DNA-free RNA was tested for purity using PCR for GAPDH genomic locus. Total RNA was tested on 2100 Bioanalyzer (Agilent Technologies) to ensure a RIN quality higher than 9 and quantified using Nanodrop. Equal amounts of total RNA (1 µg to 2.5 µg) were taken forward for library preparation and ERCC RNA Spike-in control mixes (Ambion) were added according to manufacturer guide. Ribosomal RNA was depleted using the Ribo-Zero Gold rRNA Removal module (Epicentre, Illumina) and isolated mRNA was tested for purity on 2100 Bioanalyzer (Agilent Technologies). mRNA was quantified with Qubit and equal amounts used for cDNA synthesis and 3' terminal tagging using ScriptSeq v2 RNA-seq library preparation kit (Epicentre, Illumina). The library was PCR amplified to add adaptors and barcodes. ScriptSeq v2 RNA-seq library preparation kit
GEO
GSM3610222
GEO Accession
GSM3610222