Team:Groningen/14 June 2010
From 2010.igem.org
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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. | 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. | ||
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+ | '''David & Peter''' | ||
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+ | '''Extraction of Streptomyces cellwalls''' | ||
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+ | <br> | ||
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+ | Harvesting of chaplin coated mycelium of Streptomyces with a razorblade. | ||
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+ | <br> | ||
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+ | Disruption of mycelium. | ||
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+ | <br> | ||
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+ | [[ExtractionCellwallsGR]] | ||
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+ | <br> | ||
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+ | Chaplin isolation/purification. | ||
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+ | <br> | ||
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+ | [[TFA treatmentGR]] | ||
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+ | <br> | ||
{{Team:Groningen/Footer}} | {{Team:Groningen/Footer}} |
Revision as of 12:37, 24 October 2010
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
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.
Chaplin isolation/purification.