An early whole-genome study of ethanol exposure utilized whole-embryo cultures of mouse (Liu et al. 2009). Time-mated embryos were explanted 8.25 days after conception (E8.25) into sterile bottles containing medium and grown in a rotating system for up to 48 hours. This allowed for precise ethanol exposure. Following 4 hours of pre-culture, the medium was adjusted to 88 mM (400 mg/dL) ethanol and the bottle was sealed. This dose is within the observed range but well above the mean for neonatal blood alcohol content as summarized in a recent review (Burd et al. 2012). This allowed for evaluation of the effect of blood alcohol concentrations on fetuses during the key process of neural tube closure. Samples from the cultured embryos were classified as Neural Tube Open (NTO) or Closed (NTC). DNA was extracted and immunoprecipitated with a 5mC-specific antibody, amplified, and compared with unprocessed DNA samples using Nimblegen arrays focusing on CpG islands and promoter regions. The authors categorized all available RefSeq promoters based on GC content within a defined region of 1,300 bp upstream and 500 bp downstream of
