title: GSM3610096: Mmus_shAdar2_1-2; Mus musculus; RNA-Seq
library_construction_protocol: Neurons were harvested 7 days after dissection from E16.5 brains. RNA was extracted using the PARIS kit (Ambion) followed by TURBO DNase treatment (Thermo Fisher). 250 ng of RNA was mixed with an equal amount of pooled DNA oligos designed antisense to human rRNAs in 50 bp sections in an 8 ul reaction with 2ul of 5X Hybridization buffer (500 mM Tris-HCl pH 7.4, 1  M NaCl). rRNA antisense oligos were annealed to total RNA samples for 2 minutes at 95°C, the temperature was slowly reduced to 65°C and 2U of Thermostable Hybridase H (Epicentre) and 1 ul of 10X Digestion buffer (500 mM Tris-HCl, 1 M NaCl, 200mM MgCl2) were added to make 10 ul total and reactions were incubated for 30 minutes at 65°C. rRNA-depleted RNA was then purified using Agencourt RNAClean XP beads, treated with TURBO DNase (Ambion) and then purified with RNAClean XP beads again. rRNA-depleted RNA was used as input to the KAPA RNA HyperPrep Kit (KK8540) to make RNA-seq libraries.
identifiers.namespace: GEO
identifiers.id: GSM3610096