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Apex Master Mix PCR reactions (50µl) were supplemented with MnCl2, MgCl2, dCTP, and dTTP to conduct manganese error‐prone PCR as described (Cadwell & Joyce, 1992). The mPing template was amplified with mPing TTA For and mPing TTA Rev primers (Table S3) in two rounds of mutagenic PCR to generate a library. The library was cloned into the ADE2 gene by cotransforming it with HpaI digested pWL89a plasmid into yeast. A total of 112 clones were screened for transposition to identify the mmPing20 hyperactive clone.
Copyright and License information: ©2021 The Authors. published by American Society of Plant Biologists, Society for Experimental Biology and John Wiley & Sons Ltd.
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