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Monitoring transfection efficiency by,,,green fluorescent protein (GFP) detection,,,with the Agilent 2100 bioanalyzer

This Application Note describes how the Agilent 2100 bioanalyzer and the Cell Fluorescence LabChip kit can be used to determine the efficiency of transfection of mammalian cells using green fluorescent protein (GFP) as a reporter molecule. Transfection of CHO-K1 cells with expression vectors encoding GFP from two different sources, and optimization of transfection conditions for both plasmids were performed. Histogram quality and the percent transfected cells determined, based on the low number of fluorescent cells counted with the microfluidic system, is in good agreement with data obtained with a conventional flow cytometer. Detailed protocols and reagent recommendations for analyzing transfection reactions are given. High reproducibility of chip results, low cell consumption and ease-of-use are advantages the compact Agilent 2100 bioanalyzer offers for monitoring transfection efficiency using GFP.
The Agilent 2100 bioanalyzer was introduced by Agilent Technologies as the first commercially available lab-on-a-chip analysis system for the life science laboratory using LabChipproducts, developed by Caliper Technologies Corp. Chip-based approaches for a variety of separation-based techniques have been introduced, addressing DNA, RNA, and protein separations. The Agilent 2100 bioanalyzer is capable of two-color fluorescence detection and runs disposable microfluidic glass chips. The application presented here is based on the controlled movement of cells through these channels by pressure-driven flow. Cells are hydrodynamically focused in the channels before passing the fluorescence detector in single file. Each chip accommodates several samples and data acquisition of all samples is fully automated while analysis allows for user-specific settings. Spec
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