title: GSM1976953: PAL-S; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using miRNeasy kit (Qiagen) from either freshly purified splenic B cells, obtained after gradient separation using Lympholyte Mammal (Cedarlane) followed by CD43 negative selection (Miltenyi Biotec), or from purified splenic B cells stimulated in vitro (106 cells m/L) for 48 hours in RMPI 1640 medium (Lonza) supplemented with 10% fetal calf serum (Lonza) and with 5µg/mL LPS (Cayla Invivogen), separated on a Lympholyte Mammal gradient. For each genotype (wt, 3’PALD/D, hs3b-4D/D, 3’RRD/D), RNA sample was prepared from a pool of equivalent numbers of purified B cells from 4 animals 1 µg of total RNA were depleted from ribosomal RNA, ligated, reverse transcribed and amplified (14 cycles) with the reagents from the truseq stranded total RNA kit (illumina). Libraries were quantified with the Bioanalyzer DNA 1000 Kit (Agilent). 4nM of libraries were then diluted and denatured according to the Illumina recommendations.  Paired-end 150-bp reads were sequenced on an Illumina Nextseq 500 sequencer.
attributes.tag: GEO Accession
attributes.value: GSM1976953