Team:Groningen/Notebook/Peter

From 2010.igem.org

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.

Pellicle: ROK + SigF

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)