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Team:Groningen/Notebook/Peter
From 2010.igem.org
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3x appr. 15 mg samples + ROK, DegU, SigF medium (+ 1 control normal medium) <br> | 3x appr. 15 mg samples + ROK, DegU, SigF medium (+ 1 control normal medium) <br> | ||
3x appr. 10 mg samples + ROK, DegU, SigF medium (+ 1control normal medium) <br> | 3x appr. 10 mg samples + ROK, DegU, SigF medium (+ 1control normal medium) <br> | ||
| + | |||
| + | normal medium is with 2% glucose for lysis | ||
| + | normal medium is TY for the rest | ||
<br> | <br> | ||
| + | |||
| + | 12.45: lysis at 37 degrees for 1 hour, 20 mg/ml lysozyme (control also) | ||
Revision as of 10:55, 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)
normal medium is with 2% glucose for lysis normal medium is TY for the rest
12.45: lysis at 37 degrees for 1 hour, 20 mg/ml lysozyme (control also)
