Extended Data Fig. 10: DNA detection with SPARDA. | Nature Microbiology

Extended Data Fig. 10: DNA detection with SPARDA.

From: DNA-targeting short Argonautes complex with effector proteins for collateral nuclease activity and bacterial population immunity

Extended Data Fig. 10

a, Scheme of the fluorescent ssDNA and dsDNA substrates. b, Kinetics of fluorescence increase after guide-dependent recognition of target DNA by NbaSPARDA, using dsDNA (left) and ssDNA (right) beacon substrates. The concentrations of target DNA in the samples are shown on the right. Means and standard deviations from 3 measurements. The reactions were performed with 1 µM NbaSPARDA, 1.4 µM gRNA and serial dilutions of tDNA20 at 30 °C in a reaction buffer containing 20 mM HEPES pH 7.5, 5 mM MnCl2, 5 µg/ml BSA, 100 mM NaCl, 2 mM DTT. c, Scheme of the assay with PCR amplification followed by exonuclease digestion of the non-target DNA strand. d, Sensitivity of the assay without amplification (top, molar concentrations of the ssDNA target) and with PCR amplification (bottom, number of molecules in the sample before amplification). Means and standard deviations from 3 replicates. The data correspond to the 150 and 100 min time points from Fig. 4b and 4e, respectively.

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