IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/09/06
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2x 1% Agarose gels made, one 100ml and one 60ml. Loaded the 16S sequenced PCRs in the 100ml gel with a 1Kb+ ladder, and run at 110mV for 50 mintues fig 1.
Loaded the GUS PCR in the 60ml gel, and run for 50 minutes at 110mVfig 2.
4x 500ml SFM made. More streak purifications and lawns carried out.
Of the 10 16S lanes, 8 ran, and GUS showed enough product to gel extract, using the QIAqick kit. Excised fragments from the gels were dissolved in Buffer QG, incubated and centrifuged through a spin colum with isopropanol at 13,000rpm for one minute. This was washed with Buffer PR and eluted in sterile water, and stored in the freezer.
The 16S will be prepped and sent off for sequencing.
The biobrick K1041000 was compared to Bba_J04450 by performing a restriction digest of both with NdeI and EcoRI+PstI and running the samples on an Agarose gel.