TAE: Difference between revisions

Revision as of 12:35, 5 August 2011

TAE is a commonly used buffer for making and running DNA agarose gels. It offers a few advantages and disadvantages compared to TBE buffer:

TAE buffer provides optimal resolution of fragments >4 kb in length, while TBE provides better resolution for 0.1 to 3 kb fragments.
TAE is significantly cheaper to make
TAE stocks can be 50X concentrated and therefore take up less space than 10X concentrated TBE stock
TBE offers a higher resolution and has a higher buffering capacity at greater temperatures induced by relatively higher voltages
TBE can negatively influence the yield of DNA after recovery from a gel when using glass-based protocols. Yield does not seem to be influenced when using silica-based methods.
The bromide ion in TBE is a powerful inhibitor of enzyme activity. This can prevent degradation of nucleic acid, but can also interfere with subsequent experiments like cloning DNA into a vector.

Add dH₂O up to one litre.

To make 1x TAE from 50X TAE stock, dilute 20ml of stock into 980 ml of deionised water. and

@@ Line 1: / Line 1: @@
-==Ingredients==
+TAE is a commonly used buffer for making and running DNA agarose gels.
-* [[Tris]]
+It offers a few advantages and disadvantages compared to TBE buffer:
-* Acetate
+*TAE buffer provides optimal resolution of fragments >4 kb in length, while TBE provides better resolution for 0.1 to 3 kb fragments.
-* [[EDTA]]
+* TAE is significantly cheaper to make
+* TAE stocks can be 50X concentrated and therefore take up less space than 10X concentrated TBE stock
+* TBE offers a higher resolution and has a higher buffering capacity at greater temperatures induced by relatively higher voltages
+* TBE can negatively influence the yield of DNA after recovery from a gel when using glass-based protocols. Yield does not seem to be influenced when using silica-based methods.
+* The bromide ion in TBE is a powerful inhibitor of enzyme activity. This can prevent degradation of nucleic acid, but can also interfere with subsequent experiments like cloning DNA into a vector.
-To make 1x TAE from 10X TAE, dilute 400ml of 10X TAE into 3.6 L of deionized water and store in a 4 L jug.
+==Ingredients for one litre 50X stock==
+* [[Tris-base]]: 242 g
+* Acetate (100% acetic acid): 57.1 ml
+* [[EDTA]]: 100 ml 0.5M sodium EDTA
+Add dH<sub>2</sub>O up to one litre.
+To make 1x TAE from 50X TAE stock, dilute 20ml of stock into 980 ml of deionised water.
+[[Category:Material]] and [[Category:Agarose gel electrophoresis]]