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 Citations (Scopus)

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
StatePublished - Dec 1 2004

Fingerprint

vortex tubes
Dynamic mechanical analysis
Amplification
polymerase chain reaction
Vortex flow
DNA
deoxyribonucleic acid
Polymerase chain reaction
cycles
bacteriophages
Denaturation
Bacteriophages
biopolymer denaturation
elongation
Elongation
assembly
fragments
Annealing
Kinetics
annealing

ASJC Scopus subject areas

  • Instrumentation

Cite this

Ranque-Hilsch vortex tube thermocycler for fast DMA amplification and real-time optical detection. / Ebmeier, Ryan J.; Whitney, Scott E.; Sarkar, Amitabha; Nelson, Michael; Padhye, Nisha V.; Gogos, George; Viljoen, Hendrik J.

In: Review of Scientific Instruments, Vol. 75, No. 12, 01.12.2004, p. 5356-5359.

Research output: Contribution to journalArticle

Ebmeier, Ryan J. ; Whitney, Scott E. ; Sarkar, Amitabha ; Nelson, Michael ; Padhye, Nisha V. ; Gogos, George ; Viljoen, Hendrik J. / Ranque-Hilsch vortex tube thermocycler for fast DMA amplification and real-time optical detection. In: Review of Scientific Instruments. 2004 ; Vol. 75, No. 12. pp. 5356-5359.
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