title: Imprinting RNAseq
design: Imprinting RNAseq
library_construction_protocol: All mice used in this study were housed in the animal facility at the Stowers Institute for Medical Research (SIMR) and handled according to SIMR and National Institutes of Health (NIH) guidelines. All procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of SIMR. H19-DMR flDMR/flDMR mice on B6 background were kindly provided by Dr. Marisa S. Bartolomei (University of Pennsylvania Perelman School of Medicine). Conditional mutant Igf1rfl/+ was kindly provided by Thomas L Clemens (Center for Musculoskeletal Research, Johns Hopkins Medicine, 601 North Caroline Street, Baltimore, MD 21287). Interferon inducible Mx1-Cre or tamoxifen inducible Scl-Cre mouse strains were used to delete the floxed H19-DMR and Igf1r. For Mx1-Cre activation, 250ug of pIpC was injected intraperitoneal every other day for 14 days at 5 weeks of age. For Scl-CreER activation, 2mg tamoxifen dissolved in 0.1 ml of corn oil was injected intraperitoneal every day for 5 days.  Illumina TruSeq RNA Sample Prep Kit (Catalog #: FC-122-1001) The RNA-sequencing library was prepared from approximately 200 ng of total RNA (mH19dDMR/+  Igf1r-/- , mH19dDMR/+Igf1rf-/- and mH19flDMR/+) for each sample.  The fragment size in the generated library ranged from 220 to 500 bps with a peak at 280 bps.
attributes.tag: Experimental Factor: genotype
attributes.value: WT
attributes.tag: Experimental Factor: cell type
attributes.value: MPP (Lin- Sca1+ cKit+ CD34+ Flk2+)