Chemical and Biomolecular Engineering Research and Publications


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This article was originally published in "Review of Scientific Instruments, Volume 75, Number 12. [DOI: 10.1063/1.1819411] © Copyright (year) American Institute of Physics. This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics. The following article appeared in Review of Scientific Instruments, Volume 75, Number 12. and may be found at AIP web site.


An innovative polymerase chain reaction (PCR) thermocycler capable of performing real-time optical detection is described below. This device utilizes the Ranque–Hilsch vortex tube in a system to efficiently and rapidly cycle three 20 mL samples between the denaturation, annealing, and elon-gation temperatures. The reaction progress is displayed real-time by measuring the size of a fluo-rescent signal emitted by SYBR green/double-stranded DNA complexes. This device can produce significant reaction yields with very small amounts of initial DNA, for example, it can amplify 0.25 fg (,5 copies) of a 96 bp bacteriophage l-DNA fragment 2.731011-fold by performing 45 cycles in less than 12 min. The optical threshold (150% of the baseline intensity) was passed 8 min into the reac-tion at cycle 34. Besides direct applications, the speed and sensitivity of this device enables it to be used as a scientific instrument for basic studies such as PCR assembly and polymerase kinetics.