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This feature, often offered with laboratory and information management systems (LIMS), is implemented for reactions, samples and analyses.
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For acetylated reactions, samples were processed as above, except that the 500 µL of supernatant (following sonication and pelleting) was acetylated according to manufacturer's protocol prior to dilution in EIA buffer and further processing.
For deglycosylation reactions, samples denatured in protein loading buffer were treated with 500 units of PNGase F (NEB) in 1× G7 Reaction Buffer and 1% NP40 for 1.5 hours at 37°C.
After adding reaction buffer, 10%NP40andnd 3 μl of PNGase F or water (for control reaction), samples were incubated at 37°C for 2 hr.
For the binding reaction, samples were incubated at 30°C for 10 min or the times indicated.
Note that, for 24-h-reaction samples, the hexagonal NaYF4 diffraction peaks were so intensive that the cubic NaGdF4 peaks were almost not observed (Fig. 2e).
For all reactions, DNA samples from the cagA-positive ATCC43504 and J99 H. pylori strains were used as positive controls.
Signals of RT-LAMP were detected within 50 min of the reaction for samples with 400 copies of viral genomes.
For all reactions, each sample was loaded in triplicate and SYBR green was used as the fluorescent dye.
Although 30 min was used for the H7N9 RT-LAMP reactions, most of the amplification reactions for clinical samples could be finished within 25 min.
A liquid handler was used to collect samples for reaction analysis.
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