MOESM8 of Recapitulation of gametic DNA methylation and its post-fertilization maintenance with reassembled DNA elements at the mouse Igf2/H19 locus
2020-01-15T05:13:02Z (GMT) by
Additional file 8: Figure S8. Monoallelic gene expression pattern is recapitulated in LCb118 knock-in mice. Gene expression analysis of the endo-LCb118 (A) or endo-LCb (B) embryos. In order to distinguish parental origin of the alleles by using SNPs between inbred mouse strains, endo-LCb118 or -LCb hetero-knock-in mice (H19 ICR/LCb118 or LCb; C57BL/6 J [B6] background) were mated with wild-type mice (H19 ICR/H19 ICR; JF1/Msf [JF1]), and offspring was obtained. Total RNA was prepared from livers of E12.5 embryos. Igf2 and H19 gene transcripts were amplified by RT-PCR (within logarithmic amplification range) with α-32P-labeled dCTP, followed by BstUI or Cac8I digestion, respectively. Parental origin of transcripts was discriminated by allele-specific restriction sites. The sites were also introduced into primer sequence so that complete digestion of PCR products can be concomitantly monitored. Gapdh gene transcript was analyzed as control.