Illumina NovaSeq 6000 paired end sequencing; RNAseq of E14.5 fetal liver isolated from wild-type control, Rps19R67∆/R67∆, Rps19R67∆/R67∆ Trp53−/− and Trp53−/− mutant embryos.
RNAseq of E14.5 fetal liver isolated from wild-type control, Rps19R67∆/R67∆, Rps19R67∆/R67∆ Trp53−/− and Trp53−/− mutant embryos.
Fetal liver from embryos from timed crossings of mating pairs of appropriate genotypes were harvested 14 days after detection of plugs (E14.5). Pregnant females were euthanized and they embryos extracted in ice-cold PBS. Fetal livers were collected in PBS and dissolved by pipetting and small aliquot was used for preparation of RNA in Trizol reagent within 1 hour after extraction of mouse embryos. Total RNA from freshly harvested fetal livers was isolated with Trizol reagent following manufacturer's protocol. 1µg of DNase I-treated RNA isolated from treated cells was used for poly-A mRNA isolation with the NEBNext Poly(A) mRNA Magnetic Isolation Module. Paired-end libraries were prepared using the NEBNext Ultra II Directional RNA Library Kit with NEBNext multiplex index oligos for Illumina (Dual Index Primers Set 1) (New England Biolabs).