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. Author manuscript; available in PMC: 2021 Apr 1.
Published in final edited form as: FASEB J. 2020 Feb 26;34(4):4956–4969. doi: 10.1096/fj.201902649R

Table 1-.

Site-directed mutagenesis primers

Mutations Forward Primer Reverse Primer
BTC, M44S TGCCTGATCGTGGTGTCCGTGGTGTTCATCATC GATGATGAACACCACGGACACCACGATCAGGCA
BTC, I52S TTCATCATCCTGGTGTCCGGCGTGTGCACCTGC GCAGGTGCACACGCCGGACACCAGGATGATGAA
BTC1, M44S CTGATCGTGGTGTCCGTGGTGTTCATC GATGAACACCACGGACACCACGATCAG
BTC1, I52S ATCATCCCCGTGTCCGGCGTGTGCACC GGTGCACACGCCGGACACGGGGATGAT
iR2-JMD1–1 V406E, Q409D TGTTCTTCAGCACCAGATCGGTGGTCTCGTGCTGGGCAAAGCC GGCTTTGCCCAGCACGAGACCACCGATCTGGTGCTGAAGAACA
iR2-JMD1–2 A403S, L410S GTGGGCTTTTCCCAGCACGAGACCACCGATTCCGTGCTGAAG CTTCAGCACGGAATCGGTGGTCTCGTGCTGGGAAAAGCCCAC
iR2-JMD1–3 A403S, V406E, Q409D, L410S GTGGGCTTTTCCCAGCACGTTACCACCCAGTCCGTGCTGAAG CTTCAGCACGGACTGGGTGGTAACGTGCTGGGAAAAGCCCAC
ADAM17 L659P GATTTCATTGACCAGCCTAGCATCAACACTTTTG CAAAAGTGTTGATGCTAGGCTGGTCAATGAAATC
ADAM17 6A.1 (first mutagenesis step) GATTTCATTGACCAGCTGGCCGCCGCCACTTTTGGGAAGTTTCTG CAGAAACTTCCCAAAAGTGGCGGCGGCCAGCTGGTCAATGAAATC
ADAM17 6A.2 (second mutagenesis step) CAGAAACTTCCCAAAAGTGGCGGCGGCCAGCTGGTCAATGAAATC GTTATCTGCCAGAAACTTGGCGGCGGCGGCGGCGGCCAGCTGGTC