Fluorescent melting curve analysis compatible with a flowing polymerase chain reactor

Tara M. Dalton; David J. Kinahan; Mark R. Davies

doi:10.1115/imece2005-80954

Fluorescent melting curve analysis compatible with a flowing polymerase chain reactor

Tara M. Dalton, David J. Kinahan, Mark R. Davies

University of Limerick

Research output: Contribution to journal › Conference article › peer-review

Abstract

A primary tool for analysing PCR product is the Fluorescent Melting Curve Analysis (FMCA). The temperature at which a double helix DNA strand denatures depends both on its length and base pair composition. Accurate measurement of this melting temperature using fluorescence allows estimations be made regarding DNA product length and composition. Current progress in development of PCR thermal cyclers has been primarily aimed at micro-channel based flowing devices. This paper addresses the challenges associated with performing FMCA analysis which is compatible with the output from a flowing PCR thermocycler. Two PCR products of significantly different lengths and base pair composition are compared using space domain FMCA. Results allow for differentiation of the PCR product, and compare favourably with results from a commercial thermal cycler. The successful application of FMCA within a channel shows its potential for use in high throughput flow based total analysis systems (μTAS).

Original language	English
Pages (from-to)	31-36
Number of pages	6
Journal	Advances in Bioengineering, BED
Volume	57
DOIs	https://doi.org/10.1115/imece2005-80954
Publication status	Published - 2005
Event	2005 ASME International Mechanical Engineering Congress and Exposition, IMECE 2005 - Orlando, United States Duration: 5 Nov 2005 → 11 Nov 2005

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10.1115/imece2005-80954

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title = "Fluorescent melting curve analysis compatible with a flowing polymerase chain reactor",

abstract = "A primary tool for analysing PCR product is the Fluorescent Melting Curve Analysis (FMCA). The temperature at which a double helix DNA strand denatures depends both on its length and base pair composition. Accurate measurement of this melting temperature using fluorescence allows estimations be made regarding DNA product length and composition. Current progress in development of PCR thermal cyclers has been primarily aimed at micro-channel based flowing devices. This paper addresses the challenges associated with performing FMCA analysis which is compatible with the output from a flowing PCR thermocycler. Two PCR products of significantly different lengths and base pair composition are compared using space domain FMCA. Results allow for differentiation of the PCR product, and compare favourably with results from a commercial thermal cycler. The successful application of FMCA within a channel shows its potential for use in high throughput flow based total analysis systems (μTAS).",

author = "Dalton, {Tara M.} and Kinahan, {David J.} and Davies, {Mark R.}",

year = "2005",

doi = "10.1115/imece2005-80954",

language = "English",

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pages = "31--36",

journal = "Advances in Bioengineering, BED",

issn = "0360-9960",

note = "2005 ASME International Mechanical Engineering Congress and Exposition, IMECE 2005 ; Conference date: 05-11-2005 Through 11-11-2005",

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T1 - Fluorescent melting curve analysis compatible with a flowing polymerase chain reactor

AU - Dalton, Tara M.

AU - Kinahan, David J.

AU - Davies, Mark R.

PY - 2005

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N2 - A primary tool for analysing PCR product is the Fluorescent Melting Curve Analysis (FMCA). The temperature at which a double helix DNA strand denatures depends both on its length and base pair composition. Accurate measurement of this melting temperature using fluorescence allows estimations be made regarding DNA product length and composition. Current progress in development of PCR thermal cyclers has been primarily aimed at micro-channel based flowing devices. This paper addresses the challenges associated with performing FMCA analysis which is compatible with the output from a flowing PCR thermocycler. Two PCR products of significantly different lengths and base pair composition are compared using space domain FMCA. Results allow for differentiation of the PCR product, and compare favourably with results from a commercial thermal cycler. The successful application of FMCA within a channel shows its potential for use in high throughput flow based total analysis systems (μTAS).

AB - A primary tool for analysing PCR product is the Fluorescent Melting Curve Analysis (FMCA). The temperature at which a double helix DNA strand denatures depends both on its length and base pair composition. Accurate measurement of this melting temperature using fluorescence allows estimations be made regarding DNA product length and composition. Current progress in development of PCR thermal cyclers has been primarily aimed at micro-channel based flowing devices. This paper addresses the challenges associated with performing FMCA analysis which is compatible with the output from a flowing PCR thermocycler. Two PCR products of significantly different lengths and base pair composition are compared using space domain FMCA. Results allow for differentiation of the PCR product, and compare favourably with results from a commercial thermal cycler. The successful application of FMCA within a channel shows its potential for use in high throughput flow based total analysis systems (μTAS).

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DO - 10.1115/imece2005-80954

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JO - Advances in Bioengineering, BED

JF - Advances in Bioengineering, BED

T2 - 2005 ASME International Mechanical Engineering Congress and Exposition, IMECE 2005

Y2 - 5 November 2005 through 11 November 2005

ER -

Fluorescent melting curve analysis compatible with a flowing polymerase chain reactor

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