Mike Barnkob:Protocols/Cloning/Solutions

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Solutions

Basic solutions for cloning work.

0.5M EDTA solution

1. Stir 186.1g disodium ethylenediamine tetraacetate (EDTA) into 800 ml of distilled water.
2. Stir the solution vigorously using a magnetic stirrer.
3. Add NaOH solution to adjust the pH to 8.0.
4. Dilute the solution to 1 L with distilled water.
5. Filter the solution through a 0.5 micron filter.
6. Sterilize in an autoclave

TBE buffer

Creates 10x TBE buffer solution

1. Mix the following in flask:
   • 108g Tris Base (Trizma base)
   • 55g Boric acid
   • 40 ml 0.5M EDTA solution (pH 8.0)
2. Store at room temperature.

Dilute 100ml into 900 dH20 for a 1x solution. Stable for 6 months at room temperature.

References

• http://chemistry.about.com/od/labrecipes/a/Edta-Solution.htm
• http://en.wikipedia.org/wiki/TBE_buffer