EGFP (enhanced green fluorescent protein) mRNA can express an enhanced green fluorescence once has been transfected into cells, which is a direct-detection reporter mRNA, used as control to study transfection and expression in mammalian cells by various fluorescence-based assays. You can observe the fluorescence at 509 nm.
Our product had already been modified with ARCA with a high efficient co-transcriptional capping method, which means most of mRNA has a Cap 0 structure. It is a more mature mRNA than mRNA with no capping and maintain a more stable state which lead to higher expressing efficiency in transfected mammalian cells. ARCA obtain the ability only inserting in the proper orientation, results in forming mRNAs that can be translated twice efficiently as those initiated with mCAP.
Please dissolve it on ice and protect from RNase carefully. Avoid repeated freeze/thaw cycles as possible. Don’t vortex. Upon first use, centrifuge the tube softly and aliquot it into several single use portions. Use RNase-free reagents and materials with appropriate RNase-free technique. Don’t add to the media with serum unless mixing with a transfection reagent.
Shipping Condition
Evaluation sample solution : ship with dry ice.
Protocol
Cell line
HEK293T
Tissue culture plate
48-well tissue culture plate
The plating cell volume
50,000
Cell density before transfection
cell fusion degree 50%-60%
Transfection conditions
After culturing for 17 hours, transfection (1μg mRNA+2μL lipofectamine 2000) in serum- and antibiotic-free medium, 4 hours later, change to complete medium to return to normal growth environment and perform fluorescence photography after 24 hours.
RNA amount and transfection reagent
1μg mRNA and 2μL lipofectamine 2000 are prepared according to the instructions
Transfection efficiency
above 90% transfection efficiency
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