Application and preparation of TRIS buffer in nucleic acid agarose electrophoresis

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Tris buffer is an integral part of nucleic acid agarose electrophoresis, and most running buffers use Tris buffer. The two main buffers used in nucleic acid agarose electrophoresis are TBE (Tris borate/EDTA) and TAE (Tris acetate/EDTA).

TRIS powder

Although there are some differences in the resolution of different forms of DNA and their mobility during electrophoresis, Tris buffers and derivatization solutions are often used interchangeably. The borate ions in TBE inhibit many enzymes, and some enzyme-mediated downstream manipulations may not work without some type of DNA purification after electrophoresis. So in most DNA labs, TAE buffers are more suitable for everyday use.

Two ways to make Tris buffer solutions

1. Prepare Tris base and Tris HCl solutions of desired concentrations by adding an aliquot of one solution (usually Tris HCl) to the other (usually Tris base) while monitoring pH until obtaining Correct pH.

2. Dissolve an appropriate amount of Tris powder in water, adjust the pH with HCl, then make up the buffer to the desired volume, this method will not change the ionic strength if there is no overshoot when adjusting the pH. The ionic strength changes once overshoot and readjustment with NaOH, or after pH adjustment with Tris HCl and NaOH.

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