GSM2816822: WT_HZ2186; Mus musculus; RNA-Seq
We performed RNA extraction and purification from the cerebellum of male mice 8 to 9 weeks of age (three biological replicates of wild type and Mecp2-null) using the Aurum™ Total RNA Fatty and Fibrous Tissue Kit (Bio-Rad 7326830) per the manufacturer's instructions. Genomic DNA was eliminated using an on-column DNase digestion step. RNA quality was assessed using the Agilent 2100 Bioanalyzer system prior to library preparation for deep sequencing or use of the total RNA for Nanostring nCounter quantification. RNA library preparation was performed using the cerebellar RNA from 3 wildtype and 3 Mecp2-null mice. We used the Illumina TruSeq Stranded mRNA library preparation protocol (publication number 15031047, rev. E) and 250ng of total RNA was used to make each library, and ERCC RNA Spike-In Mix 1 or 2 (Thermo Fisher 4456740) was added to the samples prior to library preparation according to the manufacturers instructions. Samples were paired end sequenced (PE100) on a HiSeq2500 in high-output mode using the v4 sequencing kit (Illumina; SBS: FC-401-4003, paired-end flow cell: PE-401-4001).
GEO Accession
GSM2816822