GSM2412138: WT-S1; Mus musculus; miRNA-Seq
CD43- splenic B-cells from wt, Em-deficient and 3'RR-deficient mice were cultured in conventional growth medium alone or with 5mg/ml of LPS for 2 days. Cells from two mice were pooled. Cellular RNA was then trisol-extracted. 1 µg of total RNA were ligated, reverse transcribed and amplified (17 cycles) with the reagents from the NextFlex small RNAseq kit V3 (Bioo scientific). Amplified libraries were quantified with the Bioanalyzer High Sensitivity DNA Kit (Agilent), pooled and size-selected from 140 nt to 160 nt with the LabChip XT DNA 300 Assay Kit (Caliper Lifesciences). Libraries were then sequenced on an Illumina Nextseq 500 Mid Flowcell with 75pb reads.