miniaturized spatial temperature gradient capillary electrophoresis system with radiative heating and automated sample introduction for DNA mutation detection

• Published in: Electrophoresis 2010-09, Vol.31 (18), p.3137-3143
• Main Authors: Xu, Zhang-Run, Li, Qi, Fan, Xiao-Feng, Zhang, Hui-Dan, Fang, Jin
• Format: Article
• Language: English
• Subjects: Automated sample introduction; Automation, Laboratory; DNA - analysis; DNA - chemistry; DNA - genetics; DNA mutation; DNA Mutational Analysis - instrumentation; DNA Mutational Analysis - methods; Electrophoresis, Capillary - instrumentation; Electrophoresis, Capillary - methods; Genes, ras - genetics; Hot Temperature; Humans; Miniaturization; Miniaturization - instrumentation; Reproducibility of Results; Spatial temperature gradient CE
• ISSN: 0173-0835; 1522-2683; 1522-2683
• DOI: 10.1002/elps.201000205

A miniaturized spatial temperature gradient CE system with automated sample introduction for DNA mutation detection was established. Continuous electrokinetic sample injection was achieved by combining an automated slotted-vial array sample introduction device to the spatial temperature gradient CE system. The temperature gradient was produced by a radiative heating system with a single graphite block heater, and the stability of the temperature gradient was investigated. The temperature variation of each measure point was 0.12-0.21% RSD (n=7) within 6 h. A 14-cm Teflon AF-coated silica capillary was used both as the separation channel and as the liquid-core waveguide tube of fluorescence signal. Under a temperature gradient from 54.8 to 59.5°C, a low range control mutation standard (209 bp) was separated within 4 min with only 5.6 nL sample consumption. Automated continuous sample introducing and changing were realized with a carryover of 3.3%. Utility of the system was further demonstrated by detecting K-ras gene mutations in paraffin tissue sections from two colorectal cancer patients.