Tag: TBE electrophoresis buffer

TBE (Tris-Borate-EDTA) Electrophoresis Buffer Calculator

The TBE (Tris-Borate-EDTA) electrophoresis buffer contains Tris base, Boric acid, EDTA and has pH 8.3. Usually the stock TBE electrophoresis…

Migration of Bromophenol Blue and Xylene Cyanol in Agarose Gel Running in TBE or TAE Electrophoresis Buffer

Bromophenol blue and Xylene cyanol are the two most commonly used tracking dyes for the analysis of DNA on agarose gel electrophoresis. These negatively charged dyes not only help in monitoring the progress of agarose gel electrophoresis but also allow easy monitoring of sample loading process onto the wells of agarose gel. Their position in relation to DNA fragments is an important information that helps to determine how far a gel must be run without accidentally letting the DNA fragments of interest run out of the gel and at the same time ensuring good resolution among different size DNA fragments.

Preparation of 5X TBE Electrophoresis Buffer

TBE is a very common electrophoresis buffer for DNA agarose gel electrophoresis. It is prepared as a 5X stock solution that can be diluted to 1x or 0.5x working solution for DNA agarose gel electrophoresis. The 5X concentrated solution contains 0.445 M Tris borate and 0.01 M EDTA (pH 8.2 – 8.4).

A 5X stock solution is prepared by dissolving 54 g Tris base, 27.5 g Boric acid, and 20 ml of 0.5 M EDTA in water to a final volume of 1000 ml. The solution is sterilized by autoclaving and is stored at room temperature.