Thermal calibration

Abstract

The present invention relates to the use of one or more amplicons as temperature calibrators. In some embodiments, the calibrators may be used to calibrate the temperature of a microfluidic channel in which amplification and/or melt analysis is performed. In some embodiments, the amplicons may be genomic, ultra conserved elements and/or synthetic. The amplicon(s) may have a known or expected melt temperature(s). The calibrators may be added to primers of study or may follow or lead the primers of study in the channel. The amplicon(s) may be amplified and melted, and the temperature(s) at which the amplicon(s) melted may be determined. The measured temperature(s) may be compared to the known temperature(s) at which the amplicon(s) was expected to melt. The difference(s) between the measured and expected temperatures may be used to calibrate/adjust one or more temperature control elements used to control and/or detect the temperature of the channel.

What is claimed is: 
     
         1 . A thermal calibrant comprising RFCAL-100. 
     
     
         2 . A thermal calibrant comprising RFCAL-200. 
     
     
         3 . The thermal calibrant of  claim 2 , further comprising RFCAL-100. 
     
     
         4 . Reagents comprising a test sample containing genomic DNA, at least a pair of primers for one or more nucleic acids of interest and a pair of primers for a calibrant. 
     
     
         5 . The reagents of  claim 4 , wherein the calibrant is contained within the genomic DNA. 
     
     
         6 . The reagents of  claim 5 , wherein the calibrant is a sequence from an ultra conserved element. 
     
     
         7 . The reagents of  claim 6 , wherein the sequence from the ultra conserved element is a sequence from a human chromosome 17 UCE sequence. 
     
     
         8 . The reagents of  claim 4 , wherein the calibrant is a synthetic sequence. 
     
     
         9 . A thermal calibrant system comprising a first amplicon having a known melting temperature and a second amplicon having a second known melting temperature.