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Hydroxy naphthol blue (HNB) was more effective to detect the LAMP product compared to the Real-time LAMP and turbidity assay for its simple and distinct detection.
On the other hand, probes unrelated to the sequence of the amplified LAMP product are not taken up by the LAMP-PEI complex because they are not hybridized to the LAMP product.
The LAMP product was verified by sequencing a 150 bp portion.
The hybridization of AuPr to a complementary sequence on the LAMP product made the gold resistant to high salt concentration.
Next, the LAMP product was hybridized at 63 °C for 5 min with an optimal FITC-labeled probe that was designed specifically for the LAMP amplicons.
In contrast, in the PPase-containing group, the variance in Tm values was smaller and there was no overlap in the Tm values obtained for all fungi tested: the LAMP product of each fungus had a specific Tm value, and the average Tm value increased as the GC% of the starting dumbbell-like structure increased.
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Moreover, we were able to visualize the fluorescence in the precipitate even if the specific LAMP product was 0.2 μg.
Thus, the calibration was generated by plotting the DNA LAMP product concentration against initial DNA copy numbers, as shown in Figure 6.
As shown in Figure 2c, the nucleic acid concentration of LAMP product from LAMP zones without paper inside kept decreasing through the whole experimental period.
Several methods for the detection of LAMP product were also performed to show the diverse way of detection which may be used in different settings depending on the user's infrastructure and resource.
The percentage of probes captured in the precipitate of the PEI-LAMP product complex was calculated according to the following formula based on the obtained results.
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