BpiI (BbsI) (10 U/µL)

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	LRP6 CRISPR-Cas9 targeting construct was generated by ligating the annealing product of the primers listed in Supplementary Table 1 to pX330 (obtained from Addgene #48139) digested with BbsI (Thermo Scientific).

Protocol tips
LRP6 CRISPR-Cas9 targeting construct was generated by ligating the annealing product of the primers listed in Supplementary Table 1 to pX330 (obtained from Addgene #48139) digested with BbsI (Thermo Scientific).

Publication protocol

Myc-LRP6 was a gift of C. Niehrs, Institute of Molecular Biology, Mainz, Germany. LRP5-Myc-His was a gift of G. Bu, Mayo Clinic, Jacksonville, United States. mWnt1 was a gift of H. Clevers, Hubrecht Institute, The Netherlands. LRP6 CRISPR-Cas9 targeting construct was generated by ligating the annealing product of the primers listed in Supplementary Table 1 to pX330 (obtained from Addgene #48139) digested with BbsI (Thermo Scientific). Single clones were established and genotyped. For genotyping, genomic DNA was isolated using QIAamp DNA micro kit (Qiagen). Primers for the PCR amplification using GoTaq Flexi DNA polymerase (Promega) are listed in Supplementary Table 1. Products were cloned into pGEM-T Easy vector system I (Promega) and subsequently sequenced using a T7 sequencing primer.

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