Fluorescent intensities of passive reference dye and a fluorescein amidite reporter dye are acquired and measured at the end of PCR cycles.
In this paper, a fully differential, low-power, passive reference voltage sharing SAR ADC architecture is presented, along with its theoretical analysis and test results.
Maybe it's due to a heightened awareness of climate change and environmental conditions, or a passive reference to cloud computing, or maybe the contemporary art world just has its proverbial head in the clouds these days.
ROX, a passive reference dye, was used for signal normalization across the plate.
The fluorescence intensity was normalized to the rhodamine derivative ROX as a passive reference label, which was present in the buffer solution.
The ROX passive reference dye was used to normalize for non-PCR related fluorescence signal variation (e.g., well and pipetting variability).
Approximately 10 ng of this DNA was analyzed in a 20 µl SYBR green PCR reaction containing 1X Brilliant SYBR Green QPCR Master Mix (Stratagene), 30 nM ROX passive reference dye, and 300 nM each primer.
Transcript levels were determined using a SYBR Green based system containing ROX as passive reference and run on a Stratagene Mx3000p machine (95°C, 10 min; 40 cycles of 95°C, 15 sec; 60°C, 60 sec).
In brief, PCR amplification was performed in a 384 well plate with a 20-µl reaction mixture containing 300 nm of each primer, 200 nm probe, 200 nm dNTP in 1x real time PCR buffer and passive reference (ROX) fluorochrome.
One microliter of cDNA from normal or TCC samples, respectively, was amplified in a PCR reaction with 2x Brilliant SYBR® Green QPCR Master Mix (containing 2.5 mM MgCl2), 300 nM of each primer and 30 µM Rox passive reference dye in a final volume of 20 µl.
The reactions contained the passive reference dye, ROX.
