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Additional resources for ARRL UHF Microwave Projects [Vol 2]
9. The gel should only be fixed (usually in a combination of acid and alcohol) after the gel has been imaged since the use of ethanol can interfere with the fluorescent properties of the CyDyes. The gel should not be fixed if Western blotting will be Table 2 The colors related to the different CyDyes used in a minimal labeling experiment CyDye Reagent color Laser excitation Emission fluorescence Cy2 Yellow Blue Green Cy3 Red Green Orange Cy5 Blue Red Red 18 P. Beckett performed. If the gels need to be stored prior to scanning, then they can be kept under SDS running buffer in a lighttight container at 4°C.
4. Friedman DB, Hill S, Keller JW, Merchant NB, Levy SE, Coffey RJ, Caprioli RM (2004) Proteome analysis of human colon cancer by twodimensional difference gel electrophoresis and mass spectrometry. Proteomics. 4(3):793–811. Chapter 4 Assessing Signal-to-Noise in Quantitative Proteomics: Multivariate Statistical Analysis in DIGE Experiments David B. Friedman Abstract All quantitative proteomics experiments measure variation between samples. When performing large-scale experiments that involve multiple conditions or treatments, the experimental design should include the appropriate number of individual biological replicates from each condition to enable the distinction between a relevant biological signal from technical noise.
Some important aspects of the analysis procedure are outlined below. 1. Cropping Even when working with pre-defined scan areas, raw gel images generated by the scan device contain non-essential information that should be deleted prior to performing image analysis. e. cutting the section of interest from the raw image file and pasting it as a new image into a new file. Within one MFA experiment, all cropped image sections have to be exactly of the same size and should cover the same gel section.