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. 2016 Jul 5;7:1009. doi: 10.3389/fmicb.2016.01009

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Copyright © 2016 Salvadori, Junges, Morrison and Petersen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Workflow for markerless genome editing. Amplicon design and construction (I to III) are prepared according to the desired mutation, exemplified in this diagram as a gene deletion (marked in red). This step is followed by a highly efficient (>10%) method for natural transformation (IV). Two dozen colonies recovered from the transformation step are screened with specific primers that amplify fragments with different sizes in the mutant when compared to the parent (V). Isolation of colonies with only the new recombined chromosomal DNA (VI).