Microscopy & Microtechniques

Using A CCD-Based Imager To Scan Pace Gels Provides A Quick And Sensitive Method Of Analysis

Mar 23 2007

Author: Florence Goubet, PhD, Paul Dupree, PhD, Alexandra Godlee and Paru Oatey, DPhil on behalf of Unassigned Independent Article

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Analysis of polysaccharides can be problematic because of the need for complex methods and/or expensive equipment such as chromatography and mass spectrometry [1,2], NMR [3], capillary electrophoresis [4], and Fourier transform infrared spectroscopy [3,5]. These techniques are ideal for a particular polysaccharide or application, but they have limitations, such as the need for pure compounds or relatively large sample quantity. Recently, a method known as PACE (polysaccharide analysis using carbohydrate gel electrophoresis) has been described [6,7]. This can be used in a high throughput manner and is applicable to small quantities of material. PACE relies on derivatising reducing ends of sugars and oligosaccharides with a fluorophore label, followed by polyacrylamide gel electrophoresis. To obtain sensitive detection of fluorescent signals on PACE gels generally requires laser based scanners which can be expensive; limited to fluorescence applications; or require large amounts of bench space. To overcome these difficulties, this article describes how CCD-based technology can be used to detect the fluorescent signal produced on a PACE gel and the sensitivity this type of system can achieve.

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