TAE: Difference between revisions
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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:Buffers]] | |||
[[Category:Agarose gel electrophoresis]] |
Latest revision as of 17:42, 20 March 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.