GSM8324361: retina, Samd7 KO, rep2; Mus musculus; RNA-Seq
After euthineasia with carbon dioxide, the retina was isolated from the eye and stored in RNAlater (invitrogen) until processing. RNA extraction from one retina per replicate using Rneasy mini with on column RNAse-free Dnase treatment (Qiagen). Lysis was performed in 450 ul of lysis buffer using a pestle gun. 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 15 cycles using primers incorporating unique dual index tags.