Team:TU Darmstadt/Protocols/Ammonium sulfate precipitation
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== Ammonium sulfate precipitation == | == Ammonium sulfate precipitation == | ||
+ | === About === | ||
+ | Ammonium sulfate precipitation is a method used to purify [https://2012.igem.org/Team:TU_Darmstadt/Protocols#Protein proteins] by altering their solubility. It is a specific case of a gerneral technique known as [https://2012.igem.org/Team:TU_Darmstadt/Protocols/Salting_Out salting out]. | ||
=== Materials === | === Materials === | ||
==== Equipment ==== | ==== Equipment ==== |
Revision as of 15:31, 16 September 2012
Contents |
Ammonium sulfate precipitation
About
Ammonium sulfate precipitation is a method used to purify proteins 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