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Gene Knockout Analysis of MoDcl1 and MoDcl2 with Flanking Probes and Hygromycin Resistance

This study explores the knockout of the MoDcl1 and MoDcl2 genes in mutant strains, detailing the use of gene-specific probes and flanking sequences for successful identification and validation. We employ a ladder method to characterize the knockout (KO) and ectopic integration of the hygromycin resistance marker (hygR). The analysis includes specific lengths of flanks (2833 bp) for accurate gene targeting and assessments of potential mutations. Results will contribute to understanding gene function and resistance mechanisms in these models.

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Gene Knockout Analysis of MoDcl1 and MoDcl2 with Flanking Probes and Hygromycin Resistance

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  1. (A) WT  MoDcl1 knockout  MoDcl2 in  MoDcl1 mutant (B) MoDcl1 MoDcl2/MoDcl1 5’ Flank 3’ Flank Gene Ladder KO Ect Ladder KO KOEct Probe BAR 5’ Flank 3’ Flank HindIII HindIII 1429 bp Flank out Hyg/BAR-specific (C) 1 2 3 4 5 6 7 8 9 10 11 (D) 1 2 3 4 Gene-specific Probe hygR 5’ Flank 3’ Flank HindIII PstI 2833 bp

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