Tris-borate EDTA , preparation

Standard research-grade agarose is usually sufficient, but special agaroses may be used for specific applications (e.g. high-resolution gels). The electrophoresis buffer is usually prepared and stored as a 10 x concentrated stock. The most commonly used buffer is Tris-borate-EDTA (TBE), or alternatively, one may use Tris-acetate-EDTA (TAE) buffer ... 

Tris-borate-EDTA (TBE) buffer (5x) 445 mM Tris, 445 mM borate, 10 mM EDTA, prepared with RNase-ffee H2O. 

Photopolymerizable formulations of gels from acrylamide and bis-acrylamide powder were prepared by combining monomer, crosslinker, and 7.2 g urea (a denaturant additive commonly used in DNA electrophoresis) in deionized water to make a total volume 16 mL. A 4 mL volume of lOx Tris-Borate-EDTA (TBE) electrophoresis buffer (Extended Range TBE Buffer Bio-Rad Laboratories, Hercules, CA) was then added to bring the total volume to 20 mL (6 M final urea concentration). Immediately before each experiment, 3 pL of a freshly prepared (10% w/v) aqueous ammonium persulfate (APS) solution was added, along with 2.5 pL of a (0.4% w/v) aqueous riboflavin solution, to serve as a photoinitiator... 

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