Cloning and mutagenesis primers | |||
---|---|---|---|
Primer | Sequence | Purpose | |
RNF43 InFusion F | GTTTAAACTTAAGCTTATGAGTGGTGGCCACCAG | RNF43-BirA*-HA into pcDNA4 | |
RNF43 InFusion R | AAACGGGCCCTCTAGACTATGCGTAATCCGGTACA | ||
del87–186 | ATTAATGCAGTCCCACGAGCTGAAGGAGCCCC | Site-directed mutagenesis, deletion of the PA domain | |
del87–186-antisense | GGGGCTCCTTCAGCTCGTGGGACTGCATTAAT | ||
Quantitative Polymerase Chain Reaction | |||
GENE | Forward primer seq. | Reverse primer seq. | Product size |
B2M | CACCCCCACTGAAAAAGATG | ATATTAAAAAGCAAGCAAGCAGAA | 167 |
GAPDH | GACAGTCAGCCGCATCTTCT | TTAAAAGCAGCCCTGGTGAC | 127 |
RNF43 | TTTCCTGCCTCCATGAGTTC | CAGGGACTGGGAAAATGAATC | 116 |
ZNRF3 | GCTTTCTTCGTCGTGGTCTC | GCCTGTTCATGGAATTCTGAC | 91 |
CTNNB1 | AAAGCGGCTGTTAGTCACTGG | CGAGTCATTGCATACTGTCCAT | 215 |
AXIN2 | TACACTCCTTATTGGGCGATCA | TTGGCTACTCGTAAAGTTTTGGT | 151 |
CRISPR/Cas9 Genotyping | |||
Primer | Sequence | Purpose | |
RNF43 F | AGGTCTTTTACAAGTAGATAATAGCAAGG | CRISRP/Cas9 RNF43 RFLP analysis | |
RNF43 R | GCTGCAACCCTTTCCCA | ||
ZNRF3 F | TTCTCAGCCTTGCTCTTCTG | CRISRP/Cas9 ZNRF3 RFLP analysis | |
ZNRF3 R | TCACCCTGGCCTTGAGTC |