title: GSM5231641: tKO26b/a1/a2 d15 neural cell miR-26a1/a2/b knockout CCC22; Mus musculus; RNA-Seq
library_construction_protocol: Total RNAs from two independent clones of WT and tKO26b/a1/a2 cell lines and two   biological replicates of NCs (day 15) were isolated using peqGold RNA pure   (PeqLab). rRNA was removed using the Ribo-Zero rRNA Removal Kit (Illumina). cDNA library preparation using NEB Next Ultra Directional RNA Library  Prep Kit for Illumina (New England Biolabs). For small RNA sequencing total RNA was isolated and a 3´barcoded adapter was ligated usig truncated T4 RNA ligase (WT_R1_A: 5´-GTAGAGTGACTGTGGAATTCTC-3`; WT_R1B:5´-GTAGAGTCTGTGTGGAATTCTC-3´; WT_R1C:5´-GTCCGCTGACTGTGGAATTCTC-3´; WT_R1D:  5´- GTCCGCTCTGTGTGGAATTCTC-3´; CCC_22A:  5´-GTAGAGGCGATATGGAATTCTC-3´;  CCC_22B: 5´-GTCCGCTCATAGTGGAATTCTC-3´; CCC_55A: 5´-GTAGAGTCATAGTGGAATTCTC-3´; CCC_55B:5´-GTCCGCGCGATATGGAATTCTC-3´). Ligated RNAs were pooled and size selected. Small RNA cDNA libraries were generated  using SuperscriptIII (Invitrogen) and sequenced  by Illumina Sequencing.  (Hafner et al., 2010)