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Team:Bielefeld-Germany/Results/S-Layer/Guide/5
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| + | [[Image:IGEM-Bielefeld2011-Pump.JPG|400px|thumb|center|A Milipore Pellicon XL 300 membrane actuated with a SciLog TANDEM 1081 peristaltic pump.]] | ||
Centrifugation does not remove all of the cell debris so a tangential flow ultrafiltration step with a 300 kDa membrane follows. 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 stucked after some time...). | Centrifugation does not remove all of the cell debris so a tangential flow ultrafiltration step with a 300 kDa membrane follows. 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 stucked after some time...). | ||
| - | With the cleared lysate the histidine affinity tag comes into play - [[Team:Bielefeld-Germany/Results/S-Layer/Guide/ | + | With the cleared lysate the histidine affinity tag comes into play - [[Team:Bielefeld-Germany/Results/S-Layer/Guide/6 | want to know how?]]. |
Revision as of 21:13, 27 October 2011
Filtration of the cell lysate
Centrifugation does not remove all of the cell debris so a tangential flow ultrafiltration step with a 300 kDa membrane follows. 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 stucked after some time...).
With the cleared lysate the histidine affinity tag comes into play - want to know how?.
