IGEM:University of Chicago/2009/Notebook/Paraoxon Biosensor/2009/08/03

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August 3, 2009
Concentrations of DNA + dilution: template-RFP: 7.42 ng/μL diluted to 74 pg/μL template-mcherry: 326 ng/μL diluted to 65.0 pg/μL
 * Received primers for changing GFP to RFP/mcherry!
 * Diluted primers to 100 μM in TE buffer (spin down, add TE)
 * Let rest for 1-2 hours at room temperature

For both mcherry and RFPTurbo beads: 2μL 5' primer: 5μL 3' primer: 5μL template: 2μL ddH2O: 36μL Total: 50μL
 * PCR

Program name - RFP Incubation: 95°C  5 minutes Separation: 95°C  30 seconds Annealing: 58°C   30 seconds Extension: 72°C   1.5 minutes Repeat x30
 * Fragments should be 1kbp long


 * Control 1 - No primers
 * Control 2 - no DNA (use RFP turbo primers and template)

2 beads 5' primers: 5μL 3' primers: 5μL temp 2μL ddH2O 46: 38μL


 * Ran a gel at 150V - should see band at ~750bp for mcherry and RFP turbo
 * gel will be inserted here!
 * very thick band seen at ~750
 * will do restriction digest/ligation tomorrow