GSM2753447: Embryo_E9 [P455]; Mus musculus; RNA-Seq
Trunk from the anterior limit of the forelimb bud to the tailbud were isolated, aiming to exclude all extra-embryonic, head, cervical and heart tissues. Isolates were stored in RNAlater at -80°C. Extraction was performed using the RNAeasy micro kit (Qiagen) according to manufacturer's instructions. On-column DNase I treatment was performed during 20 min at room temperature to ensure pure, DNA-free RNA sample. TruSeq Stranded Total RNA with Ribo-Zero kit from Illumina was used for the library preparation with 200 ng of total RNA as input. Library molarity and quality was assessed with  Qubit and Tapestation using a DNA High sensitivity chip (Agilent Technologies). RNA-seq. The 4 libraries were pooled and loaded at 8 pM for clustering on a Paired-end Illumina Flow cell. Reads of 100 bases were generated using the TruSeq SBS HS v3 chemistry on an Illumina HiSeq 2500 sequencer. The preparation of libraries and sequencing were performed by the genomic platform of the University of Geneva.