Team:TU Darmstadt/Protocols/Ammonium sulfate precipitation

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== Ammonium sulfate precipitation ==
== Ammonium sulfate precipitation ==
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=== About ===
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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

  1. add 1/10 volume of 7M ammonium acetate to DNA solution
  2. add 3 volumes 97% EtOH to DNA solution
  3. precipitate for 2 hours at -20°C
  4. centrifuge for 30 min at 13000 rpm and 4°C
  5. discard supernatant
  6. dry pellet for 30 min at 50°C
  7. solve pellet in 30 µL water

Notes

  • Important: Cooling must be maintained at all times.

References

  • Praktikumsskript Biotechnologie 2012 TU Darmstadt
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