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Starting concentrations for Chloroauric acid and BSA solutions: Chloroauric acid --> 17.0 mg in 10 mL of water --> 4.3 mM BSA --> 10.3 mg in 10 mL of water --> 15.4 uM Used these stocks to make solutions for synthesizing nanoparticles. Use the same final concentrations that you used on the 7th. Also, use the same buffers that your groups were assigned to. Tomorrow we will all use water for the synthesis. I will give you directions for how we are conducting this experiment at the beginning of class.
Make LB/agar plates with antibiotic (will describe in class)
Place sterile tube and your DNA (post DpnI reaction) in ice bucket for 15 minutes while thawing and cells.
Add 5μL of your DNA and 40μL of cells to the bottom of the tube.
Incubate on ice for 30 minutes
Heat shock your DNA/Cells at 42C for 30seconds
Incubate on ice for 5 minutes
Add 250μL of SOC media (I will supply this)
Incubate in shaker at 37°C for 1 hour
Spread 100μL of cells on your LB/agar plate
Store plate (inverted) in 37°C oven over
NOTE:wrong calculations were used were corrected for tomorrows lab