"Our analysis shows that a simple linear correction for a 40 degrees Celsius reference temperature identified errors between minus 3 to 8 degrees Celsius for three previously published sets of calibration equations derived at approximately 23 degrees Celsius," says lead researcher Shah.
To address the problem, the NIST team developed mathematical methods to correct for the shift experienced when the reference temperature changes. This allowed the researchers to create generalized calibration equations that can be applied to any reference temperature.
Microfluidic DNA amplification (production of numerous copies of DNA from a tiny sample) by the polymerase chain reaction (PCR) is one procedure that could benefit from the new NIST calculations, Shah says. "PCR requires a microfluidic device to be cycled through temperatures at three different zones starting around 65 degrees Celsius, so a useful dye intensity-to-temperature ratio would have to be based on that temperature and not a reference point of 23 degrees Celsius," she explains.
|Contact: Michael E. Newman|
National Institute of Standards and Technology (NIST)