Fig. 3: Experimental validation of primers and gRNAs for Streptococcus pyogenes in an RPA/CRISPR–Cas12a assay. | Communications Biology

Fig. 3: Experimental validation of primers and gRNAs for Streptococcus pyogenes in an RPA/CRISPR–Cas12a assay.

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

Fig. 3

a Distribution of guide RNAs across the S. pyogenes ATCC 19615 genome (RefSeq assembly accession GCF_000743015.1). b Fluorescence profiles of guide RNAs across 35 min for the S. pyogenes ATCC 19615 strain. Data represent the average fluorescence values of three experimental replicates. Error bars represent the standard error of the mean (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 S. pyogenes gDNA, with NTC as the negative control. Data represent the normalized fluorescence at the assay endpoint for four replicates. Samples were compared to the NTC using a one-sided t-test.

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