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Tuesday, May 14, 2013

50X TAE Buffer Preparation - Tris Acetic EDTA Buffer


TAE Buffer- Tris Acetic EDTA Buffer

TAE Buffer consists of Tris Base, Acetic acid and EDTA, a buffer which is most commonly used as running buffer for agarose gel electrophoresis in the separation of nucleic acids (DNA and RNA).

Preparation of 50X TAE Buffer Stock Solution

Chemical
Weight / Volume
Molarity
Tris Base
242 gms
2 Molar
Acetic Acid
57.1mL

0.5M EDTA pH 8.0
100 mL
50 milli Molar
MilliQ
up to 1000 mL


Final Concentration in 1X TAE Buffer
Tris – 40mM
EDTA - 1mM

Preparation 1X TAE From 50X TAE Stock

This can be calculated using the formula:

C1*V1 = C2*V2

C1 - Initial Concentration
V1 - Initial Volume
C2 - Final Concentration
V2- Final Volume

For Example, you want to make 1X TAE Buffer from 50X TAE Buffer Stock then how much 50X TAE is required,
50 * V1 = 1*300
V1 = 300 / 50 = 6mL
So to make 300mL of 1X TAE Buffer you should take 6mL of 50X TAE Stock solution and make it up to 300mL using MilliQ water (6mL of 50X TAE + 294mL of Water).