GSM6359080: ATM_WT_12wks_4; Mus musculus; RNA-Seq
GSM6359080
RNA was extracted from cells or tissue using RNeasy Plus Mini or Micro kit (Qiagen) RNA sequencing libraries were prepared using the SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Clontech/Takara) from 10ng of total RNA. This protocol includes a first step of RNA fragmentation using a proprietary fragmentation mix at 94°C. The time of incubation was set up for all samples at 4 min, based on the RNA quality, and according to the manufacturer's recommendations. After fragmentation, indexed cDNA synthesis and amplification were performed followed with a ribodepletion step using probes targeting mammalian rRNAs. PCR amplification was finally achieved on ribodepleted cDNAs, using 12 cycles estimated in accordance to the input quantity of total RNA. Sequencing was carried out using a pair-end 2 x 100 bp mode on the NovaSeq 6000 system (Illumina), targeting between 10 and 15M clusters per sample