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Strategy for amplification and mutagenesis of the M. pneumoniae glpK gene (MPN050 [6]). The positions of the opal codons in the wild-type glpK gene (indicated by a W followed by the number corresponding to the amino acid) and the position and orientation of the external and mutagenic oligonucleotides are shown. The annealing site of each oligonucleotide is indicated by an arrow. Oligonucleotides bearing an A→G transition are depicted by crossed arrows.
