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Issue 9, 2007
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DNA mutation detection with chip-based temperature gradient capillary electrophoresis using a slantwise radiative heating system

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Abstract

A simple and robust chip-based temperature gradient capillary electrophoresis (TGCE) system was developed for DNA mutation/single-nucleotide polymorphism (SNP) analysis using a radiative heating system. Reproducible, stable and uniform temperature gradients were established along a 3 cm length of the electrophoretic separation channel using a single thermostated aluminium heater plate. The heater was slightly slanted relative to the plane of the glass chip at 0.2–1.3° by inserting thin spacers between the plate and chip at one end to produce differences in radiative heating that created the temperature gradient. On-chip TGCE analyses of 4 mutant DNA model samples amplified from plasmid templates, each containing a single base substitution, with a wide range of melting temperatures, showed that mutations were successfully detected under a wide temperature gradient of 10 °C and within a short gradient region of about 3 cm (3.3 °C cm−1 gradient). The radiative heating system was able to establish stable spatial temperature gradients along short microfluidic separation channels using simple peripheral equipment and manipulation while ensuring good resolution for detecting a wide range of mutations. Effectiveness of the system was demonstrated by the successful detection of K-ras gene mutations in 6 colon cancer cell lines.

Graphical abstract: DNA mutation detection with chip-based temperature gradient capillary electrophoresis using a slantwise radiative heating system

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Publication details

The article was received on 05 Feb 2007, accepted on 07 Jun 2007 and first published on 26 Jun 2007


Article type: Paper
DOI: 10.1039/B701649B
Citation: Lab Chip, 2007,7, 1162-1170
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    DNA mutation detection with chip-based temperature gradient capillary electrophoresis using a slantwise radiative heating system

    H. Zhang, J. Zhou, Z. Xu, J. Song, J. Dai, J. Fang and Z. Fang, Lab Chip, 2007, 7, 1162
    DOI: 10.1039/B701649B

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