Difference between revisions of "IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/09/03"

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(Floor One)
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==Floor One==
 
==Floor One==
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[[Image:GEL6.png|thumb| Fig 1. Gel image of the digested pMS82. Lane 1 and 2 are 1Kb+ ladders, and lanes 3 and 4 are the digested plasmid. Lane 5 is a control of undigested pMS82. The lower band in lanes 3 and 4 is the excised neomycin gene, but the band above that is unknown.]]
 
Made 1% agarose gel.<br>
 
Made 1% agarose gel.<br>
 +
 
Filtered, concentrated and resuspended spore stocks in 20% glycerol.<br>
 
Filtered, concentrated and resuspended spore stocks in 20% glycerol.<br>
Performed agarose gel electrophoresis on NdeI and HindIII pMS82 digest products.<br>
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Performed agarose gel electrophoresis on NdeI and HindIII pMS82 digest products ''fig 1'' found that there were two bands in addition to the undigested plasmid. Unsure what the middle band could be.<br>
 +
 
 
Performed a gel extraction using a QIAquick kit on the top (undigested pMS82) and middle (unsure) bands.<br>
 
Performed a gel extraction using a QIAquick kit on the top (undigested pMS82) and middle (unsure) bands.<br>
 +
 
Set up 4 ligation reactions with the 2 extracted vectors, each with and without ligase.<br>
 
Set up 4 ligation reactions with the 2 extracted vectors, each with and without ligase.<br>
Set up overnight cultures of the 36 spore stocks that produced anti-fungal compounds in the Bioassays. A 10 ul sample of spores was inoculated in 10 ml LB and incubated at 37 °C O/N.
 
  
 +
Set up overnight cultures of the 36 spore stocks that produced anti-fungal compounds in the Bioassays (examples in ''fig 2''). A 10 ul sample of spores was inoculated in 10 ml LB and incubated at 37 °C O/N.
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[[Image:bioassays.png]]
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Fig 2. Bioassay plates showing two drops of spore stock (left is sample 38.3^-4 and right is sample 24.6^-6) which has been challenged with SNA containing ''C.albicans''.
  
  

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Floor One

Fig 1. Gel image of the digested pMS82. Lane 1 and 2 are 1Kb+ ladders, and lanes 3 and 4 are the digested plasmid. Lane 5 is a control of undigested pMS82. The lower band in lanes 3 and 4 is the excised neomycin gene, but the band above that is unknown.

Made 1% agarose gel.

Filtered, concentrated and resuspended spore stocks in 20% glycerol.

Performed agarose gel electrophoresis on NdeI and HindIII pMS82 digest products fig 1 found that there were two bands in addition to the undigested plasmid. Unsure what the middle band could be.

Performed a gel extraction using a QIAquick kit on the top (undigested pMS82) and middle (unsure) bands.

Set up 4 ligation reactions with the 2 extracted vectors, each with and without ligase.

Set up overnight cultures of the 36 spore stocks that produced anti-fungal compounds in the Bioassays (examples in fig 2). A 10 ul sample of spores was inoculated in 10 ml LB and incubated at 37 °C O/N. Bioassays.png

Fig 2. Bioassay plates showing two drops of spore stock (left is sample 38.3^-4 and right is sample 24.6^-6) which has been challenged with SNA containing C.albicans.