Team:Bielefeld-Germany/Results/S-Layer/Guide/5
From 2011.igem.org
(Difference between revisions)
Line 3: | Line 3: | ||
=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, 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...). | 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...). |
Revision as of 01:35, 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?