An ideal polymerase chain reaction (PCR) system should be capable of rapidly
amplifying a wide range of targets in both single and multiplex formats. Unfortunately,
the timescales and complexities involved in many existing technologies impose
significant limitations on achievable throughput. Buoyancy driven PCR is emerging as a
simplified version of thermally driven bio-analysis systems. Here, we demonstrate a
simplified convectively driven thermocycler capable of performing single and multiplex
PCR for amplicons ranging from 191 bp to 1.3 kb within 10 to 50 minutes using 10 to 25
µL reaction volumes. By positioning two independent thermoelectric heating elements
along the perimeter of a flow loop reactor constructed using ordinary plastic tubing, a
buoyancy-driven flow is established that continuously circulates reagents through
temperature zones associated with the PCR process. Unlike conventional benchtop
thermocyclers, this arrangement allows reactions to be performed without the need for
dynamic temperature control of inactive hardware components while maintaining
comparable product yields and requiring no modifications to standard PCR protocols.
We also provide a general correlation that can be applied to design reactor geometries
satisfying virtually any combination of reagent volume and cycling time. In addition to offering an attractive combination of cost and performance, this system is readily
adaptable for portable battery powered operation, making it feasible to perform PCRbased
assays in a broader array of settings.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/ETD-TAMU-1060 |
| Date | 15 May 2009 |
| Creators | Agrawal, Nitin |
| Contributors | Ugaz, Victor M |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | Book, Thesis, Electronic Dissertation, text |
| Format | electronic, application/pdf, born digital |
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