The exact integration site for the BptfXG023 insertion allele was confirmed by sequencing a PCR product generated by amplifying genomic DNA from a heterozygous mouse tail clipping. The DNA was amplified using primers JL511 and JL515 (see Table S3 for primer sequences). Reactions were heated for 3 min at 94°C, then cycled for 35 cycles at 30 sec at 94°C, 30 sec at 65°C, and 60 sec at 72°C, followed by one cycle for 2 min at 72°C. A ∼1.8 Kb PCR product was resolved on 1.5% agarose and purified. The product was sequenced using Big Dye v3.1 using primer JL511 to obtain BptfXG023 junction sequence.
