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Fig. 4 | Plant Methods

Fig. 4

From: Efficient mutagenesis and genotyping of maize inbreds using biolistics, multiplex CRISPR/Cas9 editing, and Indel-Selective PCR

Fig. 4

Indel-Selective PCR Assay. (A) Single base insertion/deletion (+/-N) created by CRISPR mutagenesis. To design wild type and indel allele specific forward primers (denoted by F1 & F2, respectively), the single base indel is positioned three nucleotides away (shown as -3) from the 3’ end of the primers. Reverse primer is common to assays for both wild-type and mutant alleles. (B) Illustration of 3’ end of allele-specific primer annealing only to their respective DNA template resulting in PCR amplification. (C) Schematic of the agarose gel electrophoresis analysis for wild type and mutant assay revealing the zygosity and genotype of tested DNA samples. (D) Gel image displaying PCR amplification of 503 bp DNA product for NRT1.1a and nrt1.1a mutant genotyping assays. (E), Gel image displaying PCR amplification of 338 bp DNA product for NRT1.1b and nrt1.1b mutant genotyping assays

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Plant Methods

ISSN: 1746-4811