Extended Data Fig. 4: Methodological characterization and analysis of CRISPR-Cas12a-based heme detection.
From: High-yield porphyrin production through metabolic engineering and biocatalysis
a. Fluorescence intensity of detection system in the presence of heme and its structural analogues (n = 3 biological independent detection of fluorescence intensity per condition). Fluorescence intensity was measured in the presence of heme, coproporphyrin I, coproporphyrin III, Fe-coproheme III, and protoporphyrin, with a concentration of 200 nM for heme and 2000 nM for each of the remaining porphyrin compounds. b. The linear relationship between the concentration of heme and the cleavage rate. The concentration range in which the calibration curve was linear was 10–200 nM (n = 3 biological independent replicates; R2 > 0.993). Bar graphs with error bars in a and b represent mean ± s. d. c. Analysis of the reproducibility and accuracy of heme detection method. Three different concentrations of heme samples (sample 1, 2, 3) were independently tested in six times (n = 6) using a 96-well plate. The coefficient of variation (c.v.) and sample recovery (Rec.) rate were calculated for the detection results.
