GSM8282918: wt3_dn, Bulk RNA seq; Mus musculus; RNA-Seq
Total RNA was extracted using RNeasy Micro Kit (Qiagen) per manufacturer's instructions. Samples were prepared according to library kit manufacturer's protocol (SMARTer Low Input RNA Kit, Clontech Takara Bio), indexed, pooled, and sequenced on an Illumina NovaSeq 6000. Library preparation was performed with 10 ng total RNA with a Bioanalyzer RNA integrity number score greater than 8. Double standard complementary DNA (cDNA) was prepared using the SMARTer Ultra Low RNA kit for Illumina Sequencing (Takara-Clontech) according to the manufacturer's protocol. cDNA was fragmented using a Covaris E220 sonicator using the following settings: peak incident power 18, duty factor 20%, and cycles per burst 50 for 120 s. 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 to 15 cycles using primers, incorporating unique dual index tags. Fragments were sequenced on an an Illumina NovaSeq 6000