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Fig. 2 | Plant Methods

Fig. 2

From: An optimised CRISPR/Cas9 protocol to create targeted mutations in homoeologous genes and an efficient genotyping protocol to identify edited events in wheat

Fig. 2

Schematic representation of the genotyping protocol to detect CRISPR editing events in wheat genes. Rows starting with Genome A, B and D illustrate the three homoeologous genes with the best match to TaNFXL1, with black and white boxes representing coding and non-coding exons respectively, horizontal lines introns and light grey boxes sgRNA positions. Horizontal arrows indicate the position of the homoeolog-specific PCR primers used for the first round of PCR. FAM, NED and VIC fluorescent dyes were used in a second PCR amplification to label the amplicons from the homoeologs on subgenomes A, B and D respectively. The bottom panel is a schematic representation of an electropherogram depicting possible results for non-edited (WT) and CRISPR-edited (nfxl1) homoeologs from subgenomes A, B and D

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Plant Methods

ISSN: 1746-4811