Difference between revisions of "TAE"

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Tris-Acetate buffer keeps the solution slightly basic, which keeps DNA and soluble (deprotonated, removes H<sup>+</sup>). EDTA sequesters metal ions which are used by many enzymes (particularly divalent [[Magnesium|Mg<sup>2+</sup>]]).  
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Tris-Acetate buffer keeps a solution slightly basic (approximate [[pH]] of 8.3), which keeps the DNA water soluble (the DNA is deprotonated, H<sup>+</sup> is removed). EDTA sequesters (chelates) metal ions which are used (as a co-factor) by many enzymes (particularly divalent [[Magnesium|Mg<sup>2+</sup>]] ions), some of which can degrade the DNA.  
  
 
=50X Stock Solution=
 
=50X Stock Solution=
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100 [[ml]] 0.5 [[M]] [[EDTA]]  
 
100 [[ml]] 0.5 [[M]] [[EDTA]]  
  
Add water up to a 1 [[L]] total volume, mix and give it time to dissolve.
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Add water up to a 1 [[L]] total volume, mix, and give it time to dissolve.
  
 
=1X Working Solution=
 
=1X Working Solution=

Latest revision as of 04:37, 28 July 2018

Tris-Acetate buffer keeps a solution slightly basic (approximate pH of 8.3), which keeps the DNA water soluble (the DNA is deprotonated, H+ is removed). EDTA sequesters (chelates) metal ions which are used (as a co-factor) by many enzymes (particularly divalent Mg2+ ions), some of which can degrade the DNA.

50X Stock Solution

242 g Tris

57.1 ml Acetate

100 ml 0.5 M EDTA

Add water up to a 1 L total volume, mix, and give it time to dissolve.

1X Working Solution

Add 20 ml of 50X stock to 980 ml of water.

Links

https://www.ncbi.nlm.nih.gov/pubmed/15351274

https://en.wikipedia.org/wiki/TAE_buffer

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