Difference between revisions of "TAE"

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==1X TAE==
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TAE is a commonly used buffer for making and running DNA agarose gels.
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It offers a few advantages and disadvantages compared to TBE buffer:
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*TAE buffer provides optimal resolution of fragments >4 kb in length, while TBE provides better resolution for 0.1 to 3 kb fragments.
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* TAE is significantly cheaper to make
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* TAE stocks can be 50X concentrated and therefore take up less space than 10X concentrated TBE stock
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* TBE offers a higher resolution and has a higher buffering capacity at greater temperatures induced by relatively higher voltages
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* 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.
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* 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.
  
*10X TAE
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==Ingredients for one litre 50X stock==
*Deionized water
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* [[Tris-base]]: 242 g
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* Acetate (100% acetic acid): 57.1 ml
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* [[EDTA]]: 100 ml 0.5M sodium EDTA
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Add dH<sub>2</sub>O up to one litre.
  
Dilute 400ml of 10X TAE into 3.6 L of deionized water and store in a 4 L jug.
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To make 1x TAE from 50X TAE stock, dilute 20ml of stock into 980 ml of deionised water.
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[[Category:Material]] and [[Category:Agarose gel electrophoresis]]

Revision as of 11:35, 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.

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 dH2O up to one litre.

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