GSM3391718: WT rep 3; Mus musculus; RNA-Seq
Telencephalons from E12.5 embryos were dissected, dissociated with StemPro Accutase, and passed through a 40-μm cell strainer to obtain single-cell suspensions. Cells were counted and 1.0 x 106 cells were pelleted by centrifugation. Diluted ERCC spike-in mix (1:20, Thermo Fisher Scientific 4456740) was added to cell pellets and Trizol prior to RNA extraction at a ratio of 1 μl of diluted ERCC spike-in mix per million cells. RNA was purified using a Direct-zol RNA kit (Zymo Research R2052) and eluted in 25 μl of elution buffer. Library preparation and sequencing were performed by the Genome Sequencing Facility at St. Jude Children's Research Hospital. RNA quality was checked by a 2100 Bioanalyzer RNA 6000 Nano assay (Agilent) before library generation. Libraries were prepared from 350 ng of total RNA using the TruSeq Stranded Total RNA Library Prep Kit (Illumina 20020597), then quantified using the Quant-iT PicoGreen dsDNA assay (Life Technologies). Samples were sequenced on Illumina HiSeq4000, obtaining 100 million 100-bp paired-end reads. RNAs from 5 control and 4 dKO embryos were sequenced.
GEO Accession
GSM3391718