GSM4255678: MON 1233; Mus musculus; RNA-Seq
After Ficoll separation of peripheral blood, monocytes (Gr-1+Mac-1+) were sorted from the mononuclear fraction. Total RNA was extracted using TRIzol and the Ambion PureLink RNA Mini Kit (Life Technologies, cat# 12183018), treated with DNase to remove genomic DNA using the DNAfree Kit (Life Technologies, cat# AM1906). Sequencing libraries were constructed from 100 ng of DNase-treated, total RNA using the ScriptSeqv2 Complete Kit (Illumina cat# BHMR1224). The RNA depleted for rRNA using the Ribo-Zero removal reagents and fragmented. First strand cDNA was then synthesized using a 5' tagged random hexamer and reverse transcription, followed by annealing of a 5' tagged, 3'-end blocked terminal-tagged oligo and second strand synthesis. The Di-tagged cDNA fragments were purified, barcoded, and PCR-amplified for 15 cycles.
GEO Accession
GSM4255678