GSM5013432: Periostin knockout 2; Mus musculus; RNA-Seq
RNA isolation was performed using RNeasy Spin Column method (Qiagen). RNA quality and concentrations were measured by Agilent Bioanalyzer. ds-cDNA was prepared using the SMARTer Ultra Low RNA kit for Illumina Sequencing (Takara-Clontech) per manufacturer's protocol. cDNA was fragmented using a Covaris E220 sonicator using peak incident power 18, duty factor 20%, cycles per burst 50 for 120 seconds. 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 12-15 cycles using primers incorporating unique dual index tags. Fragments were sequenced on an Illumina HiSeq 3000 using single-end reads extending 50 bases.