Illumina HiSeq 2500 paired end sequencing; RNA-Seq of Sall4 genetically modified  mouse embryonic stem cells compared to wild type controls
RNA-Seq of Sall4 genetically modified  mouse embryonic stem cells compared to wild type controls
The cells used are mouse embryonic stem cells. For RNAseq ESC lines were grown for 7-10d to obtain a healthy, fast growing culture. Then 3x105 cells were seeded to 6wells and cells were grown for 48h before isolating RNA using the AllPrep DNA/RNA kit (Qiagen). RNA was prepared from three independent cell seedings to obtain triplicate samples.  DNA was removed using the TURBO DNAfree kit (Ambion) and RNA integrity was assessed on a bioanalyzer. 4Âµg RNA were used and ERCC Spike-ins were added to prepare sequencing libraries using the ScriptSeq Complete Gold Kit (Illumina). ScriptSeq Index Primers (Illumina) were used to multiplex libraries.