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Team:Bielefeld-Germany/Results/S-Layer/Guide/5
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{{Bielefeld_2011_Header}} | {{Bielefeld_2011_Header}} | ||
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=Filtration of the cell lysate= | =Filtration of the cell lysate= | ||
| - | [[Image:IGEM-Bielefeld2011-Pump.JPG|400px|thumb| | + | [[Image:IGEM-Bielefeld2011-Pump.JPG|400px|thumb|right|A Milipore Pellicon XL 300 membrane actuated with a SciLog TANDEM 1081 peristaltic pump.]] |
| - | Centrifugation does not remove all of the cell debris | + | Centrifugation does not remove all of the cell debris, therefore a tangential flow ultrafiltration step with a 300 kDa membrane is performed. The retentate is collected. Instead of this device you can also use a simple dead-end sterile filter (0.22 µm poresize) but it is easier the way described above (the dead-end filter gets clogged after some time...). |
With the cleared lysate the histidine affinity tag comes into play - [[Team:Bielefeld-Germany/Results/S-Layer/Guide/6 | want to know how]]? | With the cleared lysate the histidine affinity tag comes into play - [[Team:Bielefeld-Germany/Results/S-Layer/Guide/6 | want to know how]]? | ||
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Latest revision as of 02:47, 29 October 2011
Filtration of the cell lysate
Centrifugation does not remove all of the cell debris, therefore a tangential flow ultrafiltration step with a 300 kDa membrane is performed. The retentate is collected. Instead of this device you can also use a simple dead-end sterile filter (0.22 µm poresize) but it is easier the way described above (the dead-end filter gets clogged after some time...).
With the cleared lysate the histidine affinity tag comes into play - want to know how?
