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FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).
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FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).
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FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).
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FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).
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FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).
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FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).

Journal: iScience

Article Title: High-throughput screening of FRET-based proteins using a hyperspectral microcapillary array

doi: 10.1016/j.isci.2026.115302

Figure Lengend Snippet: FRET constructs and ratio (A) Plasmid designs of various constructs are illustrated. The sequences of three linkers and two tandem repeats (TR) proteins used in the study are shown. (B) Representative image of E. coli cultures expressing each construct. (C) Scatterplot of fluorescence intensity obtained in the green and red channels with the plate reader. The dotted and solid circles denote two biological replicates per sample with eight technical replicates each. FRET ratios measured by HyCAP are significantly higher than those determined by the plate reader, owing to differences in the excitation and emission measurement setups. FRET ratio distributions of replicates measured using plate reader (30 replicates each) (D) and HyCAP (E).

Article Snippet: E. coli BL21 strain , New England Biolabs , C2527H.

Techniques: Construct, Plasmid Preparation, Expressing, Fluorescence