TAE
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| - | + | 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. | ||
| - | To make 1x TAE from | + | ==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. | ||
| + | |||
| + | ==See also== | ||
| + | |||
| + | For disambiguation purposes, see [[EDTA]] and [[TE]]. | ||
| + | |||
| + | |||
| + | [[Category:Material]] | ||
| + | [[Category:Agarose gel electrophoresis]] | ||
Revision as of 09:29, 13 February 2013
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.
Ingredients for one litre 50X stock
Add dH2O up to one litre.
To make 1x TAE from 50X TAE stock, dilute 20ml of stock into 980 ml of deionised water.
