Figure 4. Effect of primer coating density on the SBE reaction

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Figure 4. Effect of primer coating density on the SBE reaction Figure 4. Effect of primer coating density on the SBE reaction. Grey bar represents unspecific signal, white bar represents specific signal and black diamond represents the ratio of specific signal to unspecific signal. The error bar represents error ± standard deviation. From: New separation-free assay technique for SNPs using two-photon excitation fluorometry Nucleic Acids Res. 2004;32(13):e108. doi:10.1093/nar/gnh102 Nucleic Acids Res | Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

Figure 3. Effect of annealing temperature on the SBE reaction Figure 3. Effect of annealing temperature on the SBE reaction. Grey bar represents unspecific signal, white bar represents specific signal and black diamond represents the ratio of specific signal to unspecific signal. The error bar represents error ± standard deviation. From: New separation-free assay technique for SNPs using two-photon excitation fluorometry Nucleic Acids Res. 2004;32(13):e108. doi:10.1093/nar/gnh102 Nucleic Acids Res | Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

Figure 2. Effect of labeled nucleotide concentration on the SBE reaction. Grey bar represents unspecific signal, white bar represents specific signal and black diamond represents the ratio of specific signal to unspecific signal. The error bar represents error ± standard deviation. From: New separation-free assay technique for SNPs using two-photon excitation fluorometry Nucleic Acids Res. 2004;32(13):e108. doi:10.1093/nar/gnh102 Nucleic Acids Res | Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

Figure 1. Effect of polymerase concentration on the SBE reaction Figure 1. Effect of polymerase concentration on the SBE reaction. Grey bar represents unspecific signal, white bar represents specific signal and black diamond represents the ratio of specific signal to unspecific signal. The error bar represents error ± standard deviation. From: New separation-free assay technique for SNPs using two-photon excitation fluorometry Nucleic Acids Res. 2004;32(13):e108. doi:10.1093/nar/gnh102 Nucleic Acids Res | Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

Figure 5. Scatter plot of the signals provided by the samples representing different genotypes. Vectors of the two white data points do not exceed the vector length threshold limit set for reliable genotyping. From: New separation-free assay technique for SNPs using two-photon excitation fluorometry Nucleic Acids Res. 2004;32(13):e108. doi:10.1093/nar/gnh102 Nucleic Acids Res | Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

Figure 6. Vectors provided by the samples representing different genotypes. Vector lengths are normalized to threshold value (length of liquid fluorescence vectors). The two white data points do not exceed the threshold limit set for reliable genotyping. Vectors presented in the figure are background (liquid fluorescence) subtracted. From: New separation-free assay technique for SNPs using two-photon excitation fluorometry Nucleic Acids Res. 2004;32(13):e108. doi:10.1093/nar/gnh102 Nucleic Acids Res | Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved