title: GSM9244318: WT Meckel's cartilage sample 3; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA extraction was done with RNeasy Micro kit (Qiagen). The concentration and the purity of the total RNA extracted were measured by capillary electrophoresis using a tape Station (Agilent). The strand-specific RNA-seq libraries were prepared with the Ovation Universal RNA-Seq kit (Tecan). After a preliminary DNase digestion with a thermosensitive DNase (ArcticZymes), total RNA was reverse transcribed, and second strand of cDNA was synthetized. A fragmentation step was performed before Illumina compatible indexed adaptator ligation. The ligation was followed by the strand selection enzymatic reaction to keep the information about the orientation of the transcripts. Insert Dependent Adaptor Cleavage (InDA-C) technology was used to deplete all the cDNA corresponding to ribosomal transcripts before the PCR enrichment. To ensure that no excess of amplification was performed during the final PCR step, the number of PCR cycles applied to each sample was evaluated in a preliminary Q-PCR test using EvaGreen. An equimolar pool of the final indexed RNA-Seq libraries was sequenced on an Illumina HiSeq2500 (paired-end reads 130 bases + 130 bases) and ~50 millions paired-end reads per library were produced.
identifiers.namespace: GEO
identifiers.id: GSM9244318_r1