GSM7445302: E-cadherin knock out 3; Canis lupus familiaris; RNA-Seq
RNA was extracted using the RNeasy kit (Qiagen, Cat.No./ID:74104). Briefly cells were lysed with RLT buffer and disrupt using cell shredder. Following RNA collection following the kit protocol, RNA concentrations were obtained using nanodrop or a fluorometric Qubit RNA assay (Life Technologies, Grand Island, New York, USA). 250 ng of high quality total RNA sample ($RIN > 8$) was processed using Stranded mRNA Prep kit (Illumina) according to manufacturer instructions. Briefly, after purification of poly-A containing mRNA molecules, mRNA molecules are fragmented and reverse-transcribed using random primers. Replacement of dTTP by dUTP during the second strand synthesis will permit to achieve the strand specificity. Addition of a single A base to the cDNA is followed by ligation of Illumina adapters. Libraries were quantified by Q bit and profiles were assessed using the DNA High Sensitivity LabChip kit on an Agilent Bioanalyzer. Libraries were sequenced on an Illumina Nextseq 500 instrument using 75 base-lengths read V2 chemistry in a paired-end mode.