GSM5068471: human integrin β3-rescued, integrin β3 kockout PyMT-BO1 cells treated with DTX, rep 3 [B3KO-HB3-DTX.3]; Mus musculus; RNA-Seq
Cells were lysed and RNA extracted using the Rneasy Plus Mini Kit according to manufacturer's directions. Library preparation was performed with 1ug of total RNA. Ribosomal RNA was removed by a hybridization method using Ribo-ZERO kits (Illumina-EpiCentre). mRNA was then fragmented and reverse transcribed to yield cDNA using SuperScript III RT enzyme (Life Technologies, per manufacturer's instructions) and random hexamers. A second strand reaction was performed to yield ds-cDNA. cDNA was blunt ended, had an A base added to the 3' ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 13 cycles using primers incorporating unique index tags. Fragments were sequenced on an Illumina HiSeq3000 using single end reads extending 50 bases.