Team:Bielefeld-Germany/Results/S-Layer/Guide/5

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(Filtration of the cell lysate)
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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...).  
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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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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 07:26, 28 October 2011

Filtration of the cell lysate

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...).

With the cleared lysate the histidine affinity tag comes into play - want to know how?