TAE Buffer (10X)
Tris-Acetate-EDTA, 10x concentrated buffer for electrophoresis
TAE Buffer (10X) is an aqueous solutions of 400mM Tris, 200mM acetic acid, and 10mM EDTA, prepared with ultrapure water, and 0.2 μm filtered. TAE is the most common buffer used for agarose gel electrophoresis in the analysis of DNA samples and is recommended for high resolution of large DNA fragments (>3kb) or supercoiled DNA, however, it can also be used for non?denaturating RNA agarose electrophoresis.
Usage
The working concentration is either 1X or 0,5X. Prepare 1L TAE Buffer (1X) by mixing 100ml of the 10x concentrated buffer with 900ml of ddH2O. A 1X TAE buffer consists of 40mM Tris?acetate, 1mM EDTA at pH 8.3±0.1. TAE should be used both for the preparation of the agarose gel as well as the running buffer. Note that compared with other electrophoresis buffers, such as TBE or SGTB, TAE has
a low ionic strength and low buffering capacity and may become exhausted during extended electrophoresis (>4h). The running buffer should therefore be replaced frequently.
Storage
TAE Buffer (10X) should be stored at room temperature and is stable for up to one and a half years.
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Green DNA Stain direct
Xpert Green DNA Stain direct is a new and safe alternative to ethidium bromide (EtBr) for the visualization of DNA (double-stranded and single-stranded DNA) and RNA in agarose and polyacrylamide gels. The dye has been developed for direct loading and is compatible with both UV and Blue LED transilluminators. Xpert Green DNA Stain direct is provided as a sample loading dye. There is no need to add any dye to the gel or running buffer: just add Xpert Green DNA Stain direct to your sample and perform electrophoresis.
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