Fig. 4: SNP-Chip can differentiate between samples with and without a SOD1 SNP. | Nature Biomedical Engineering

Fig. 4: SNP-Chip can differentiate between samples with and without a SOD1 SNP.

From: Discrimination of single-point mutations in unamplified genomic DNA via Cas9 immobilized on a graphene field-effect transistor

Fig. 4

a, gRNA-CS04 was designed to target the fALS-associated H44R SNP (red) three nucleotides from the PAM (yellow). The gRNA target is highlighted (blue). b, In vitro cleavage assay with gRNA-CS04 and CS04 and WTC11 amplicons shows more efficient cleavage of CS04 than WTC11. The cleavage products are indicated by black arrowheads. The percentage cleavage was determined with ImageJ. c, I response of the Cas9-CS04 construct in the presence of CS04, WTC11 and negative control amplicons (n ≥ 5 technical replicates; F = 13.02). d, C response of the Cas9-CS04 construct in the presence of CS04, WTC11 and negative control amplicons (n ≥ 5 technical replicates; F = 16.7). Significance in c and d was determined by Tukey’s multiple comparisons test after one-way ANOVA (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001), with P < 0.05 considered significant. e, Schematic of the Cas9-CS04 construct in the presence of CS04 and WTC11 genomic DNA. f, C response in the presence of CS04 and WTC11 genomic DNA (60 ng µl−1) (t = 8.131; d.f. = 29; n ≥ 16 technical replicates). Significance was determined by two-tailed unpaired t-test (****P < 0.0001). All box and whisker plots show the minima, Q2, median, Q3 and maxima.

Back to article page