title: GSM5324703: neg2_01; Mus musculus; RNA-Seq
library_construction_protocol: On day 7, medium was removed and RNA was harvested using RNeasy Plus Micro Kit (Cat#74034). Libraries were prepared according to KAPA RNA HyperPrep Kit with RiboErase (HMR) (Cat# KK8560). Briefly, total RNA was depleted of rRNA by hybridization of complementary DNA oligonucleotides, followed by treatment with RNase H and DNase to remove rRNA duplexed on DNA and original DNA oligonucleotides. After fragmenting the RNA using heat and magnesium, the first strand cDNA was synthesized using random priming, followed by a combined second strand synthesis and A-tailing process. Libraries were amplified by PCR after the addition of adapters, and finally purified and size selected using magnetic beads. Libraries were sequenced on the Illumina NextSeq2000 following the manufacturer's protocols.