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1 2002

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Miniaturyzacja w wysokosprawnej elektroforezie kapilarnej

Mrass A., Bald E.

Wiadomości Chemiczne

2002

[Z] 56, 11-12

931-952

During the last decade high performance capillary electrophoresis (HPCE) has developed into a powerful analytical method. Despite the overwhelming advantages of this technique, it has not yet found widespread use in molecular biology labs in comparison with the slab gel electrophoresis. The main reason for the low acceptance of this method is performing separation in a single capillary which is analogous to using only one lane on a slab gel; despite the high speed of the separation, the overall throughput is still low. To comply with the current needs for fast and high throughput analysis, especially in the field of DNA sequencing, two approaches have been introduced: a multicapillary scheme called capillary array electrophoresis and a multichannel system etched into the surface of microchips. Capillary array electrophoresis (CAE) was first introduced in 1992. The search for a comprehensive instrument was achieved by several groups seeking an instrument capable of fast, automated, sensitive and most important of all, rugged operation. In CAE electrokinetic injection is the most popular method for sample loading. However adopting this mode of injection for DNA sequencing requires a thorough sample clean-up. To inject unpurified samples a so-called ‘base-stacking’ method has been described. One of the major problem in CAE is detection as it has to meet plenty of demands, among which the most important are: high speed, high sensitivity and high spatial resolution. Numerous approaches have been developed to address these challenges using either scanning or imaging technologies. Capillary electrophoresis (CE) on microchips is based upon microfabrication techniques developed in the semiconductor industry. The design of microchips for CE has undergone significant development from simple single-channel structures to increasingly complex ones capable of proceeding various analytical steps. Sensitive detection schemes are essential in microfabriacted devices in CE due to extremely small size of the detection cell. Laser induced fluorescence (LIF) is so far the most popular for this purpose. Other methods successfully coupled to microchips include mass spectrometry, electrochemical detection, Raman spectroscopy and holographic refractive index detection.