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Table 1 Forward (-F) and reverse (-R) primers used for point mutations in α ENaC (a-), and insertion of a 27 nt segment coding for a 9 residue epitope tag in β ENaC (b+) cDNA.

From: Simple and efficient site-directed mutagenesis using two single-primer reactions in parallel to generate mutants for protein structure-function studies

Primer Sequence* Length (nt)
a-R333K-F ctgtccctgatgctgAAGgcagagcagaatgacttc 36
a-R333K-R gaagtcattctgctctgcCTTcagcatcagggacag 36
a-D338A-F cgcgcagagcagaatgCcttcattcccctgctg 33
a-D338A-R cagcaggggaatgaagGcattctgctctgcgcg 33
a-R350A-F cacagtgactggggccGCggtaatggtgcacggg 34
a-R350A-R cccgtgcaccattaccGCggccccagtcactgtg 34
a-E358D-F gtgcacgggcaggatgaTcctgcctttatggatg 34
a-E358D-R catccataaaggcaggAtcatcctgcccgtgcac 34
a-K474R-F ctgggctgtttcaccaGgtgccggaagccatgc 33
a-K474R-R gcatggcttccggcacCtggtgaaacagcccag 33
a-R476K-F ctgtttcaccaagtgcAAgaagccatgcagcgtg 34
a-R476K-R cacgctgcatggcttcTTgcacttggtgaaacag 34
b+HA-F gccattgccaccaggTACCCATACGACGTCCCAGACTACGCTaacctgaacttctcc 57
b+HA-R ggagaagttcaggttAGCGTAGTCTGGGACGTCGTATGGGTAcctggtggcattggc 57
  1. * Mutated or inserted nucleotides are written in upper case characters.
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BMC Biotechnology

ISSN: 1472-6750

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