IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/31

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(Floor Two)
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==Floor One==
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To gain an idea of what strains of Streptomyces are growing on the plates, liquid cultures of the purified streaked Streptomyces plates for 16S sequencing were prepared by transferring colonies to LB (-NaCl) broth + a spring to aerate the cultures.
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Started preparing spore stocks by adding approx. 2 ml glycerol to lawn plates and using a cotton bud to shift the spores, adding more glycerol if required. The resulting liquid was then filtered using cotton wool in a 5 ml syringe. We spore stocked 32 lawns, 35 are remaining atm.
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Liquid cultures were set up for E. coli ETpuz cells - LB broth was inoculated with cultures from pAU3-45 and pMS82 plates. The pAU3-45 cells had 10 ul Apr and the pMS82 cells were inoculated with 10 ul Hyg in LB -NaCl as Hygromycin is salt-intolerant.
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500 ml SFM + 10 ml MgCl2???
==Floor Two==
==Floor Two==
A miniprep of the biobrick AntAgp+Neo was prepared and analysed by gel electrophoresis.  
A miniprep of the biobrick AntAgp+Neo was prepared and analysed by gel electrophoresis.  

Revision as of 11:50, 15 August 2013

Week Nine Main project page
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Floor One

To gain an idea of what strains of Streptomyces are growing on the plates, liquid cultures of the purified streaked Streptomyces plates for 16S sequencing were prepared by transferring colonies to LB (-NaCl) broth + a spring to aerate the cultures.

Started preparing spore stocks by adding approx. 2 ml glycerol to lawn plates and using a cotton bud to shift the spores, adding more glycerol if required. The resulting liquid was then filtered using cotton wool in a 5 ml syringe. We spore stocked 32 lawns, 35 are remaining atm.

Liquid cultures were set up for E. coli ETpuz cells - LB broth was inoculated with cultures from pAU3-45 and pMS82 plates. The pAU3-45 cells had 10 ul Apr and the pMS82 cells were inoculated with 10 ul Hyg in LB -NaCl as Hygromycin is salt-intolerant.

500 ml SFM + 10 ml MgCl2???

Floor Two

A miniprep of the biobrick AntAgp+Neo was prepared and analysed by gel electrophoresis. A colony from the E.coli BL21 plates was inoculated into LB with Kanamycin and Cholorophenicol antibiotics to perform protein over expression of the His tagged AntA σ factor.

Analysis of biobrick AntAgp+Neo using Gel electrophoresis.
Analysis of biobrick AntAgp+Neo using Gel electrophoresis.

Outreach

Finalizing posters for printing for the Forum event. Possibly, we will now have edible ants! Quality street idea as an analogy for decreasing anti-biotic discovery will be useful.

Have been contacting local radio stations (Future Radio, and radio station Matt mentioned) to see about doing an interview + promotion of the Forum event.

I have also contacted Lisa Horten from the UEA press office to see how we can publicise our project/forum event.

Possible appearance in the EDP along with Matts lab.






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