title: GSM4774992: SAM24384938: Mlkl.ko.control.LPS.8h.BMDM; Mus musculus; RNA-Seq
library_construction_protocol: Bone marrow cells from femurs and tibia of wild-type C57BL/6N mice were cultured in Roswell Park Memorial Institute (RPMI) medium  supplemented with 10% heat-inactivated fetal bovine serum, 2 mM glutamine, 50 Î¼M 2-mercaptoethanol (Gibco), 100 U/ml penicillin, and 100 Î¼g/ml streptomycin (Gibco) and 50 ng/ml  of recombinant M-CSF (R&D Systems) on non-tissue culture treated 15 cm plates. On day 5 of culture, cells were harvested and 5 million cells were re-plated onto non-tissue culture  treated 10 cm plates in fresh media (same as that described above). After 2 additional days of culture, BMDMs from 3 individual mice (each corresponding to an independent biological replicate)  were harvested by scraping. Total RNA was isolated from cells and purified using RNeasy mini kits and on-column DNAse digestion per manufacturer (Qiagen) protocol.  0.5ug used for library preparation with a TruSeq RNA Sample Preparation Kit v2 (Illumina).  The libraries were multiplexed and then sequenced on a HiSeq4000 (Illumina) to generate 30 million  single end 50 bp reads. RNA libraries were prepared for sequencing using standard Illumina protocols