Table 2.
Primers used for cloning and site-directed mutagenesis
| Primer name | Sequence (5′ to 3′) | Use |
|---|---|---|
| CDKN1B_CDS_NcoI-F | ACCCATGGGAAAGATGTCAAACGTGC | To amplify the CDKN1B CDS, adding a 5′-NcoI restriction enzyme site |
| CDKN1B_CDS-R | TTCATCAAGCAGTGATGTATCTGA | |
| CDKN1B_c.320_320delA-F | CGCAGGAGAGCCGGATGTCAGCGG | To create the c.320delA variant by site-directed mutagenesis |
| CDKN1B_c.320_320delA-R | CCGCTGACATCCGGCTCTCCTGCG | |
| CDKN1B_c.356T>C-F | CCCGGCGGCGCCTTTAACTGGGGCTCC | To create the c.356T>C variant by site-directed mutagenesis. |
| CDKN1B_c.356T>C-R | GGAGCCCCAGTTAAAGGCGCCGCCGGG | |
| CDKN1B_c.376G>C-F | GCGTGTCCTGAGAGTTAGCCGGAGCCC | To create the c.376G>C variant by site-directed mutagenesis |
| CDKN1B_c.376G>C-R | GGGCTCCGGCTAACTCTCAGGACACGC | |
| CDKN1B_c.407A>G-F | GGACCCAAAGACTGGTCCGTCGGACAGCC | To create the c.407A>G variant by site-directed mutagenesis |
| CDKN1B_c.407A>G-R | GGCTGTCCGACGGACCAGTCTTTGGGTCC | |
| RHOA_CDS_NcoI-F | GATTCGTTGCCTGAGCCATGG | To amplify the RHOA CDS, adding a 5′-NcoI restriction enzyme site |
| RHOA_CDS-R | TTAACCGCATAAGGGCTGTG | |
| HA_to_Flag_F | GGAGGTACTCACGATGGATTATAAGGATGACG | To create the Flag by site-directed mutagenesis |
| HA_to_Flag_R | CGTCATCCTTATAATCCATCGTGAGTACCTCC | |
| pSF_delHA-F | GGAGGTACTCACGATGGATTATGCGGATGACG | To delete the HA tag by site-directed mutagenesis |
| pSF_delHA-R | CGTCATCCGCATAATCCATCGTGAGTACCTCC | |
| CMV-F | CGCAAATGGGCGGTAGGCGTG | To verify inserts in plasmids by Sanger sequencing |
| R3 | AGCTGAAGGTACGCTGTATC |