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Savitzky-Golay smoothing with a window size of 50 was performed, followed by peak finding using a local maximum method with two local points.
Savitzky-Golay smoothing with a window size of 50 was first performed on spectral data followed with peak finding using a local maximum method with two local points.
A 3-D kymograph was generated by projecting each frame [a 3-D (xyz) image] to a 2-D (xy) image using the maximum method with the xy-t data visualized in the 3-D kymograph.
The threshold cycle (crossing point) in which the fluorescence rises significantly above background level was determined by a second derivate maximum method with the use of the LightCycler™ quantification software.
PCR conditions were 50°C for 2 min, 95°C for 10 min, 45 cycles of 95°C for 10 s and 65°C for 30 s, then 45°C for 5 min. Cycle threshold (Ct) values were calculated by using the second derivative maximum method with the y-axis at F1/F3 (LightCycler software version 3.5).
Standard curves describing the PCR efficiencies of the target (AdipoR1 and AdipoR2) and the reference genes (β-actin) were created from a dilution series of the calibrator cDNA (HL60 cDNA induced with phorbol myristyl acetate -PMA-), using the Second Derivative Maximum Method with the Arithmetic baseline adjustment for the determination of the various crossing points.
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The second derivative maximum method was used with the LightCycler software to automatically determine the crossing point for individual samples.
The Second Derivative Maximum Method was used with the LightCycler software to automatically determine the crossing point for individual samples as previously described [ 1].
To compare with the maximum method, the differences between their lifetimes are plotted in Figure 4.
Figure 1 Circle phylogenetic tree of 459 DHQases, constructed with Maximum Likelihood method with 1000 bootstrap replications.
The concatenate sequences of RED and TEF-1α genes were analyzed by the maximum likelihood method with 1000 bootstrap replications.
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