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Fig. 3 | BMC Microbiology

Fig. 3

From: Standardization of an LNA-based TaqMan assay qPCR analysis for Aspiculuris tetraptera DNA in mouse faeces

Fig. 3

Development of direct PCR using faeces preparations in mice. a The indicated amounts of healthy mouse faeces were diluted with TE buffer, and the final 1 ng/μl concentration of synthesized A. tetraptera DNA was added to the dilutions. The DNA preparations were amplified using the optimal PCR experimental conditions, and the PCR products were observed, as described in the “Materials and methods” section. b The DNA preparations were treated with heat at 95 °C for 5 min after dilution, as shown in (a), and the PCR was performed, as described in the legend of (a). c One % BSA, 0.1% TritonX-100, or 0.1% Tween-20 was added to the DNA preparations after the dilution, as shown in (a), and then the PCR was performed, as described in the legend of (a). d Ethanol precipitation was conducted after the dilution as shown in (a), and then the PCR was performed as described in the legend of (a). e DNA preparations after ethanol precipitation were quantified using qPCR. qPCR was conducted using the LNA-based TaqMan probes and LNA-based primers as described in the “Materials and methods” section. The data represent the mean ± SE obtained from three independent experiments. f Four micrograms of faeces from healthy mice were added to the 3-fold serial dilutions of genomic DNA extracted from the faeces of A. tetraptera-infected mice (gDNA) from 13.3 ng to 0.5 ng, and subsequently precipitated by ethanol. qPCR was conducted using the LNA-based TaqMan probe and LNA-based primers as described in the “Materials and methods” section. The data of the copy numbers were converted to logarithm. The correlation coefficient indicates the relationship between the input amounts of gDNA and the detected copy numbers. The dashed lines show the 95% confidence interval. The data represent the mean ± SE obtained from three independent experiments

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