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Quantitative DNA and Protein Analysis with a Highly Uniform UV Illumination Imaging System and Fluorescent Calibration Step Tablet
Sponsored,vendor-submitted protocol   Published in BioTechniques Protocol Guide 2007 (p.35)

Ultraviolet (UV) transillumination is a ubiquitous tool in Life Science research. With few exceptions, fluorescent stains used in post electrophoresis analysis of proteins and nucleic acids have significant excitation peaks with ultraviolet (300-365 nm) light, making midrange UV the excitation source of choice for high sensitivity analysis for many fluorophores. However, quantitative analysis has been limited by the extreme lack of illumination uniformity across the surface of typical UV light boxes. To enable quantitative UV imaging, UVP has developed and characterized the FirstLight® UV Illuminator. Through use of a high density lighting system with a tuned phosphor coating, uniformity of <5% coefficient of variance (CV) across the full imaging surface has been achieved and applied to genomic and proteomic analysis.

(Click to enlarge)

The illumination uniformity enables quantitative imaging of electrophoresis gels. Combined with UVP's VisionWorks® LS acquisition and analysis software, Biospectrum® AC automated imaging system and Fluorescent Calibration Step Tablet, the system ensures:

  • Unparalleled accuracy in day-to-day quantitative imaging of protein and DNA gels.

  • Sensitivity and dynamic range that are consistent across the illumination surface.

  • Little or no reliance on uniformity correction by software that can lead to low signal data loss.

  • Calibrated gel to gel comparison.

Calibration step tablet. (Click to enlarge)

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