title: GSM6125730: Day6_KO_Rep1; Mus musculus; RNA-Seq
library_construction_protocol: B cells were harvested by trypsinization followed by removal of feeder cells through differential adherence to tissue culture dishes for 40 minutes. RNA was extracted from B cells with a miRNeasy mini kit (217004, Qiagen), quantified with a NanoDrop spectrophotometer and its quality examined in a fragment Bioanalyzer (Aligent 2100 Bioanalyzer DNA 7500 assay). Pulsed cells were harvested by trypsinization followed by removal of feeder cells through differential adherence to tissue culture dishes for 40 minutes. RNA was extracted from Pulsed cells with a miRNeasy mini kit (217004, Qiagen), quantified with a NanoDrop spectrophotometer and its quality examined in a fragment Bioanalyzer (Aligent 2100 Bioanalyzer DNA 7500 assay). Day 4 and Day 6 cells were harvested by trypsinization followed by removal of feeder cells through differential adherence to tissue culture dishes for 40 minutes. RNA was extracted from Day 4 and Day 6 cells with a miRNeasy mini kit (217004, Qiagen), quantified with a NanoDrop spectrophotometer and its quality examined in a fragment Bioanalyzer (Aligent 2100 Bioanalyzer DNA 7500 assay). For blastocysts, RNA was extracted and retro-transcribed into cDNA using a SMART-Seq v4 Ultra Low Input RNA kit (634894, Takara). RNA concentration and quality was determined as above. Libraries for RNA-sequencing were prepared as previously described (Picelli et al., 2014). For morulas, RNA was extracted and retro-transcribed into cDNA using a SMART-Seq v4 Ultra Low Input RNA kit (634894, Takara). RNA concentration and quality was determined as above. Libraries for RNA-sequencing were prepared as previously described (Picelli et al., 2014). For RNA-sequencing, libraries were prepared with a TruSeq Stranded mRNA Library Preparation Kit (Illumina) followed by single-end sequencing (50 bp) on an HiSeq2500 instrument (Illumina), obtaining at least 40 million reads per sample.