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. 2020 May 25;21(10):3727. doi: 10.3390/ijms21103727

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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(a) Map of the plasmid used to complement the uracil auxotrophy of the CRISPR-Cas9-generated L. corymbifera mutants. Arrows indicate the recognition sites of the primers used to analyze the transformants. (b) An example of the PCR analysis of the complemented mutants. A fragment of the transferred plasmid was amplified from the pyrG-complemented CBS 429.75–pyrG^cr1/1 strain using the primers pJet1.2 forward sequencing primer and LcpyrGrev. Lane M: GeneRuler 1 kb DNA Ladder (Thermo Scientific), Lane 1: CBS 429.75–pyrG^compl/1, Lane 2: CBS 429.75–pyrG^compl/2, Lane 3: CBS 429.75–pyrG^compl/3.