IGEM:The University of Sheffield/2009/Notebook/Sheffield iGEM 2010/2010/09/14

iGEM Project name 1

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Tuesday 14th September Carried out plasmid mini preps of all the successful transformations using the overnight cultures from yesterday.[[1]] Set up an overnight digestions of the promoters using the following method: Promoter digest: Plasmid - 10μL Pst1 - 2μL EcoR1 - 2μL EcoR1 buffer - 2μL BSA (x20) - 1μL H2O - 3μL Set up an overnight digestion of part BBa_J04450, using the following method: BBa_J04450 digest: Plasmid - 10μL Pst1 - 2μL EcoR1 - 2μL EcoR1 buffer - 2μL BSA (x20) - 1μL H2O - 3μL Part BBa_J04450 is a RFP contained in a pSB1C3 plasmid and is also Cm resistant. This digestion is therefore to remove the RFP so that this plasmid can be used for submission of our promoter-GFP biobricks to iGEM.