Team:Groningen/14 June 2010

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Week 24, Arend Jan

Control digestion of 4 clones of pNZ8901-bbs (bbs = biobrick sites). The original plasmid does have a XbaI, SpeI, and PstI site (which were replaced in the PCR with these sites in the correct orientation), but no EcoRI site. Therefore the control digestion was done with EcoRI. Also, HindIII was removed in the PCR but not replaced so clones should not be cut with HindIII.

- 10ul plasmid
- 1.5ul buffer EcoRI/R
- 0.5ul EcoRI/HindIII
- 3ul MQ 

Clones 1 and 2. U = undigested plasmid, E = cut with EcoRI, H = cut with HindIII

Clones 3 and 4. Use this imager in the future

EcoRI cuts in all clones except only partially in 4. HindIII does not cut in all clones (positive control would have been nice, new enzyme however). Clones 1, 2, and 3 are good for sure.

David & Peter

Extraction of Streptomyces cellwalls

Harvesting of chaplin coated mycelium of Streptomyces with a razorblade.

Disruption of mycelium.

ExtractionCellwallsGR

Chaplin isolation/purification.

TFA treatmentGR