"
Team:Bielefeld-Germany/Results/S-Layer/Guide/5
From 2011.igem.org
(Difference between revisions)
(→Filtration of the cell lysate) |
(→Filtration of the cell lysate) |
||
| Line 5: | Line 5: | ||
[[Image:IGEM-Bielefeld2011-Pump.JPG|400px|thumb|center|A Milipore Pellicon XL 300 membrane actuated with a SciLog TANDEM 1081 peristaltic pump.]] | [[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 | + | 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]]? | ||
Revision as of 18:14, 28 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?
