Ranque-Hilsch vortex tube thermocycler for fast DMA amplification and real-time optical detection

Ryan J. Ebmeier, Scott E. Whitney, Amitabha Sarkar, Michael Nelson, Nisha V. Padhye, George Gogos, Hendrik J Viljoen

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

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 μL samples between the denaturation, annealing, and elongation temperatures. The reaction progress is displayed real-time by measuring the size of a fluorescent 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 λ-DNA fragment 2.7 × 10 11-fold by performing 45 cycles in less than 12 min. The optical threshold (150% of the baseline intensity) was passed 8 min into the reaction 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.

Original languageEnglish (US)
Pages (from-to)5356-5359
Number of pages4
JournalReview of Scientific Instruments
Volume75
Issue number12
DOIs
Publication statusPublished - Dec 1 2004

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ASJC Scopus subject areas

  • Instrumentation

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