Team:TU Darmstadt/Protocols/Ammonium sulfate precipitation
From 2012.igem.org
Contents |
Ammonium acetate / Ethanol precipitation
About
Ammonium acetate precipitation is a method used to purify DNA by altering their solubility. It is a specific case of a gerneral technique known as salting out.
Materials
Equipment
- -40°C freezer
- Centrifuge (cooling cababilities required!)
- Heat block
Chemicals & consumables
- Ammonium acetate 7M
- 97% EtOH
- Eppis
- ddH2O
Procedure
- add 1/10 volume of 7M ammonium acetate to DNA solution
- add 3 volumes 97% EtOH to DNA solution
- precipitate for 2 hours at -20°C
- centrifuge for 30 min at 13000 rpm and 4°C
- discard supernatant
- dry pellet for 30 min at 50°C
- solve pellet in 30 µL water
Notes
- Important: Cooling must be maintained at all times.
References
- Praktikumsskript Biotechnologie 2012 TU Darmstadt