Taq dna polymerase

Genomic DNA was extracted from F0 mice. PCR was performed using Taq DNA polymerase (Greiner). The primer sequences were as follows: RegKO-S 5’-CCACCATCCTTAACTGGATC-3′; RegKO-AS 5’-CTAGAGTCCATGCCAAGCAC -3′. The PCR products were purified using SV Gel and PCR Clean-Up System (Promega). The purified PCR product were sequenced using BigDye Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific) through Applied Biosystems 3500xL Genetic Analyzer (Thermo Fisher Scientific).

Genomic DNA was extracted from F0, F1, and F2 mice. PCR was performed using Taq DNA polymerase (Greiner Bio-One, Kremsmünster, Austria). The primer sequences are shown in Supplementary Material, Table S1. The PCR products were purified using the Gel/PCR Extraction Kit (NIPPON Genetics, Tokyo, Japan). The purified PCR products were sequenced using the BigDye Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific) through an Applied Biosystems 3500xL Genetic Analyzer (Thermo Fisher Scientific), or restriction enzyme digestion was performed by RspRS II enzyme (Takara Bio Inc.), according to the manufacturer’s instructions. In some cases, PCR products were cloned into TA vector (Promega, Madison, WI) before sequencing.