Fig. 4: Experimental validation of primers and gRNAs for Neisseria gonorrhoeae in an RPA/CRISPR–Cas12a assay. | Communications Biology

Fig. 4: Experimental validation of primers and gRNAs for Neisseria gonorrhoeae in an RPA/CRISPR–Cas12a assay.

From: PathoGD: an integrative genomics approach to primer and guide RNA design for CRISPR-based diagnostics

Fig. 4

a Distribution of guide RNAs across the N. gonorrhoeae FA1090 genome (RefSeq assembly accession GCF_000006845.1). b Fluorescence profiles of guide RNAs across 38 min for N. gonorrhoeae FA1090 strain. Data represent the average fluorescence values of three experimental replicates. Error bars represent the SEM. The specificity of each gRNA was tested against a panel of closely related organisms and other organisms likely to be present at the site of infection. c Limit of detection of the guide RNAs using 4000, 400, and 40 copies/μL of N. gonorrhoeae gDNA, with NTC as the negative control. Data represent the normalized fluorescence at the assay endpoint for at least three experimental repeats (n = 4 for g4966, n = 3 for all other gRNA sets). Samples were compared to the NTC using a one-sided t-test.

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