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Team:Groningen/Notebook/Peter
From 2010.igem.org
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Cell wall isolation according to protocol: Cell disruption, samples are frozen. | Cell wall isolation according to protocol: Cell disruption, samples are frozen. | ||
| - | + | Culture Bascillus over night, all four strains: Rok, DegU, SigF, Sp0A | |
10/08 | 10/08 | ||
| Line 16: | Line 16: | ||
Dry-freeze over night | Dry-freeze over night | ||
| + | |||
| + | Culture Bascillus over a extra night, all four strains: Rok, DegU, SigF, Sp0A | ||
| + | |||
| + | 11/08 | ||
| + | |||
| + | Chaplin isolation: | ||
| + | |||
| + | 4x approximately 15 mg dry cell wall (180 ul TFA), | ||
| + | 4x approximately 10 mg dry cell wall (100 ul TFA), | ||
| + | 8x approximately 5 mg dry cell wall (100 ul TFA), | ||
| + | |||
| + | Cultured bascillus strains are taken out of the stove, medium will be used for degredation experiment. | ||
| + | |||
| + | |||
| + | |||
| + | DEGREDATION EXPERIMENT: | ||
| + | |||
| + | 3x appr. 5 mg samples + ROK, DegU, SigF medium (+ 1 control normal medium) | ||
| + | 3x appr. 5 mg samples + ROK, DegU, SigF lysed medium (+ 1 control lysis normal medium) | ||
| + | 3x appr. 15 mg samples + ROK, DegU, SigF medium (+ 1 control normal medium) | ||
| + | 3x appr. 10 mg samples + ROK, DegU, SigF medium (+ 1control normal medium) | ||
Revision as of 09:03, 11 August 2010
Bascillus Subtilus, easy info:
http://www.mobitec.de/int/products/bio/04_vector_sys/index.php?bac_sub.html
http://subtiwiki.uni-goettingen.de/wiki/index.php/SubtiPathways
http://subtiwiki.uni-goettingen.de/wiki/index.php/Main_Page
09/08
Cell wall isolation according to protocol: Cell disruption, samples are frozen.
Culture Bascillus over night, all four strains: Rok, DegU, SigF, Sp0A
10/08
Cell wall isolation according to protocol: Cell disruption again, continue according to protocol
Dry-freeze over night
Culture Bascillus over a extra night, all four strains: Rok, DegU, SigF, Sp0A
11/08
Chaplin isolation:
4x approximately 15 mg dry cell wall (180 ul TFA), 4x approximately 10 mg dry cell wall (100 ul TFA), 8x approximately 5 mg dry cell wall (100 ul TFA),
Cultured bascillus strains are taken out of the stove, medium will be used for degredation experiment.
DEGREDATION EXPERIMENT:
3x appr. 5 mg samples + ROK, DegU, SigF medium (+ 1 control normal medium) 3x appr. 5 mg samples + ROK, DegU, SigF lysed medium (+ 1 control lysis normal medium) 3x appr. 15 mg samples + ROK, DegU, SigF medium (+ 1 control normal medium) 3x appr. 10 mg samples + ROK, DegU, SigF medium (+ 1control normal medium)
