IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/07/23
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July 23, 2009Genomic DNA prep, culture PCR - PCR to check if GFP-Kan unsorted in correct section in the genome. - Forward primers homologous to gene. Reverse is homologous to portion inside longtine cassette. Note: will need to make more TE for satoe. 50 mM tris pH 7.5 10 mM EDTA Reverse primer, use Kan LR - 520 bp + GFP fragment - 20 mM stock from Folu - dilute to 5 uM. start with 20 mM 25 ul + 75 ul dH2O - After shaking of EtOH (step 23) will leave O/N to evaporate EtOH - PCR w/single colony. If doesn't work, will use genomic DNA Gene/culture Code 035-1 1 35-2 2 35-5 3 139-1 4 139-2 5 49-1 6 49-2 7 186-1 8 186-2 9 213-1 10 213-2 11 236-1 12 236-2 13 346-1 14 346-2 15 PCR - 1 bead - 2.5 ul primer 1 - 2.5 ul primer 2 - 19 uL dH2O - culture spot Run on cycle iGEM 6 Spot control 16 No Primers 17 (Use 35-5) End of Day
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