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Team:UC Berkeley/ProteinPurification
From 2008.igem.org
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==Proof of Concept Experiment== | ==Proof of Concept Experiment== | ||
| + | To prove the protein purification concept, a plasmid coding for a protein with a His tag is transformed into E.coli with the arabinose induced self-lysis device. This culture will be induced to lyse with arabinose, releasing the His-tagged protein. The cellular debris is pelleted by centrifuging for 30 seconds. The supernatant will be added to a new tube and Ni-NTA will be added to it. The supernatant and Ni-NTA solution is incubated at 4 degree Celcius for 4 hours. The Ni-NTA protein complex is concentrated using a magnet. The protein is eluted from Ni-NTA and the solution is ran on a gel to see whether or not protein is purified. | ||
Revision as of 08:01, 20 October 2008
Introduction
Protein purification involves a series of step to isolate the protein of interest. Purifying a protein from E.coli first includes an extraction step, which brings the protein into solution. Depending on how
Device
Proof of Concept Experiment
To prove the protein purification concept, a plasmid coding for a protein with a His tag is transformed into E.coli with the arabinose induced self-lysis device. This culture will be induced to lyse with arabinose, releasing the His-tagged protein. The cellular debris is pelleted by centrifuging for 30 seconds. The supernatant will be added to a new tube and Ni-NTA will be added to it. The supernatant and Ni-NTA solution is incubated at 4 degree Celcius for 4 hours. The Ni-NTA protein complex is concentrated using a magnet. The protein is eluted from Ni-NTA and the solution is ran on a gel to see whether or not protein is purified.
