GSM2440709: Control_siRNA4; Mus musculus; RNA-Seq
RNA was extracted from pools of 35 morula stage embryos (93 h post-hCG injection) that had been injected with either scrambled control (5 pools) or Smchd1 siRNA (5 pools).  Each pool of embryos was an independent biological replicate. RNA was extracted using the PicoPure RNA Isolation kit (Life Technologies, Grand Island, NY) with DNase digestion to remove any contaminating DNA. Aliquots of approximately half of each sample were processed through the Ovation RNA-Seq System v2 using the Ribo-SPIATM Technology (Nugen, San Carlos, CA). This was followed by fragmentation to 300 bp using a Covaris sonicator followed by a brief S1 nuclease digestion as described ((Head et al., 2011).  After purification and end repair, the cDNA was processed through the Ovation Ultralow DR Multiplex Systems 1-8 and 9-16 for end repair, barcoding and final library production. Barcoded libraries were pooled, loaded on flowcells and sequenced with Illumina HISeq 2500 in rapid run mode to generate 50 nt single end reads. The samples were loaded at 65% of optimal loading concentration, along with PhiX Control library (Illumina) added at 10% of loading concentration to increase read sequence complexity.
GEO Accession
GSM2440709