Genotyping of transgenic mice
Plcb1+/- mice in a C57BL/6J background were kindly provided by H.-S. Shin (Kim et al., 1997). Very briefly, the knockout animals were generated by the replacement of exons that encode aminoacid residues 50–82 by a Neomycin cassette that was used both for gene disruption and for positive selection.
To genotype the mice, genomic DNA from ear punches or tail tips was extracted as previously described (Gómez-Grau et al., 2017). Genotyping was carried out by PCR on extracted genomic DNA using three specific PCR primers: a sense primer, 5′-GTTAAGTCCTCAGGCAAACACC-3’, and two antisense primers, 5′-ACCTTGGGAGCTTTGGCGTG3’ and 5′-CTGACTAGGGGAGGAGTAGAAG-3’, that allowed us to amplify a 180bp Plcb1+/+ band and a 290bpPlcb1-/- band (Filis et al., 2009). Fragments were separated by 2.5% agarose gel electrophoresis. Known WT and Plcb1+/- controls were always used to validate genotyping results. The genotype of all animals used was confirmed after the cue-induced reinstatement.