title: GSM1550373: dM_LPS7; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA  from BMDMs was isolated with TRIzol reagent (Life Technologies) according to the manufacturer protocol. Ribosomal RNA was depleted using Ribo-Zero rRNA removal kit (Epicenter). RNA was fragmented using RNA fragmentation reagent (Ambion). Enzymatically 5’-pre-adenylated RNA adaptor was ligated to RNA 3’-end. cDNA was produced using primer specific to 3’-adaptor sequence. cDNA was size selected (200-300 nt) using non-denaturing gel, recovered and circularized using Circligase II (Epicenter). DNA oligo complementary to BamHI site was annealed and digested with BamHI (ThermoScientific) in order to linearize cDNA. cDNA was the PCR-amplified with primers specific for linker regions.