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

iGEM Project name 1

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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. 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.