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Molecular characterization <t>of</t> <t>T-DNA</t> integration and Cas9 expression in transformants. (A) PCR-based screening of the primary transformants. Lane M: 100 bp DNA ladder (Thermo Fisher Scientific); lanes 1–93: DNA of the recovered plants; lane WT: wild-type; lane NTC: No template control (water); lane PC: positive control (pHSE401 vector). (B) PCR analysis of the hygromycin-resistant T 1 plants. Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR. Gene expression levels were normalized using the 2 ⁻ΔΔCt method, with GhGAPDH as the internal reference gene and the wild-type sample as the calibrator. Error bars represent the standard error (SE) of three replicates.
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Molecular characterization <t>of</t> <t>T-DNA</t> integration and Cas9 expression in transformants. (A) PCR-based screening of the primary transformants. Lane M: 100 bp DNA ladder (Thermo Fisher Scientific); lanes 1–93: DNA of the recovered plants; lane WT: wild-type; lane NTC: No template control (water); lane PC: positive control (pHSE401 vector). (B) PCR analysis of the hygromycin-resistant T 1 plants. Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR. Gene expression levels were normalized using the 2 ⁻ΔΔCt method, with GhGAPDH as the internal reference gene and the wild-type sample as the calibrator. Error bars represent the standard error (SE) of three replicates.
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Molecular characterization <t>of</t> <t>T-DNA</t> integration and Cas9 expression in transformants. (A) PCR-based screening of the primary transformants. Lane M: 100 bp DNA ladder (Thermo Fisher Scientific); lanes 1–93: DNA of the recovered plants; lane WT: wild-type; lane NTC: No template control (water); lane PC: positive control (pHSE401 vector). (B) PCR analysis of the hygromycin-resistant T 1 plants. Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR. Gene expression levels were normalized using the 2 ⁻ΔΔCt method, with GhGAPDH as the internal reference gene and the wild-type sample as the calibrator. Error bars represent the standard error (SE) of three replicates.
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Molecular characterization <t>of</t> <t>T-DNA</t> integration and Cas9 expression in transformants. (A) PCR-based screening of the primary transformants. Lane M: 100 bp DNA ladder (Thermo Fisher Scientific); lanes 1–93: DNA of the recovered plants; lane WT: wild-type; lane NTC: No template control (water); lane PC: positive control (pHSE401 vector). (B) PCR analysis of the hygromycin-resistant T 1 plants. Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR. Gene expression levels were normalized using the 2 ⁻ΔΔCt method, with GhGAPDH as the internal reference gene and the wild-type sample as the calibrator. Error bars represent the standard error (SE) of three replicates.
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Molecular characterization of T-DNA integration and Cas9 expression in transformants. (A) PCR-based screening of the primary transformants. Lane M: 100 bp DNA ladder (Thermo Fisher Scientific); lanes 1–93: DNA of the recovered plants; lane WT: wild-type; lane NTC: No template control (water); lane PC: positive control (pHSE401 vector). (B) PCR analysis of the hygromycin-resistant T 1 plants. Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR. Gene expression levels were normalized using the 2 ⁻ΔΔCt method, with GhGAPDH as the internal reference gene and the wild-type sample as the calibrator. Error bars represent the standard error (SE) of three replicates.

Journal: GM Crops & Food

Article Title: CRISPR/Cas9-mediated editing of the GhJAZ2 gene improves fiber length and lint percentage in Gossypium hirsutum L

doi: 10.1080/21645698.2026.2660546

Figure Lengend Snippet: Molecular characterization of T-DNA integration and Cas9 expression in transformants. (A) PCR-based screening of the primary transformants. Lane M: 100 bp DNA ladder (Thermo Fisher Scientific); lanes 1–93: DNA of the recovered plants; lane WT: wild-type; lane NTC: No template control (water); lane PC: positive control (pHSE401 vector). (B) PCR analysis of the hygromycin-resistant T 1 plants. Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR. Gene expression levels were normalized using the 2 ⁻ΔΔCt method, with GhGAPDH as the internal reference gene and the wild-type sample as the calibrator. Error bars represent the standard error (SE) of three replicates.

Article Snippet: Lane M: 1 kb plus DNA ladder (NEB); lanes 1–15: DNA of the hygromycin-resistant T 1 plants; lane wt: wild-type; lane NTC: No template control; lane pc: positive control. (C) Relative expression of the Cas9 gene in T 0 (i) and T 1 (ii) transformants determined by RT-qPCR.

Techniques: Expressing, Control, Positive Control, Plasmid Preparation, Quantitative RT-PCR, Gene Expression