title: GSM8992927: BMDM, LPS, 24h, rep 2; Mus musculus; RNA-Seq
library_construction_protocol: BMDMs (1 × 106) were treated as indicated and RNA was purified using Direct-zol RNA Miniprep (Zymo Research Corporation, Cat# R2053). Samples were submitted to Azenta Life Sciences for standard RNA-seq processing. RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using manufacturer's instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were initially enriched with Oligod(T) beads.  Enriched  mRNAs  were fragmented for  15 minutes  at 94 °C.  Converted  the RNA  molecules  into complementary DNA (cDNA). cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles. The sequencing library was validated on the Agilent TapeStation (Agilent Technologies, Palo Alto,  CA,  USA), and  quantified  by  using  Qubit  2.0  Fluorometer  (Invitrogen,  Carlsbad,  CA)  as  well  as  by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA).