Dataset features


Application: RNA-seq analysis
Number of samples: 20
Release date: Sep 23 2017
Last update date: Nov 20 2018
Access: Public
Genes: FGF8
Dataset link Decreased Fgf8 gene dosage on gene expression

Experimental Protocol

Mouse breeding and embryo generation: The Fgf8 neo series is a 5 member series generated from a combination of the neomycin insertion into the intron between exon 2 and 3 of the Fgf8 locus and a null allele generated from loss of exon 2. Fgf8 mice were generated from the Fgf8 flp/ floxed allele originally developed by (Meyers et al., 1998). The neo cassette was maintained in the Fgf8Neo mice. Deletion constructs were developed by crossing with β-actin cre (FVB/N-Tg(ACTB-cre)2Mrt/J), to delete exons 2 and 3 from all cells. To generate the neo series, crosses were performed between mice that were heterozygous for and mice that were homozygous for the Neo (flp) allele. Genotyping was performed as in (Meyers et al., 1998). For embryos, pregnant dams were sacrificed at embryonic day (E) 10.5 based on visualization of a post-coital plug at E0.5. E10.5 embryos were dissected into PBS on ice and snap frozen at -80C. RNA was extracted in batch preps using Trizol. TapeStation analysis was used to select samples for RNAseq, all samples tested had a RIN number of between 9.0 and 10. Single end reads were performed to a read depth of 40 million reads/ sample on an illumina NextSeq 5000.










Benedikt Hallgrimsson