Today's Benchwork
- Miniprep of O/N culture
- Follow quick vacuum protocol, except:
- At step 5, added 750 μL instead of 350 μL Neutralization solution. I "corrected" this by adding an additional 200 μL Cell Lysis solution before step 6, thus increasing the total volume.
- Quantify [ pCMV-SPORT6+BSA ]
- 95μL H2O
- + 5μL miniprepped DNA from step 1
- → measure absorbance at 260 nm and 280 nm
- PCR of BSA for cloning into pTXBI
- diluted pCMV-SPORT6+BSA to 10 μg/mL
- 5 μL plasmid + 74 μL sterile H2O
- followed Basic PCR protocol
Results
[math]\displaystyle{ [CMV-SPORT6+BSA]=(0.130AU+0.028AU)\times50\frac{\frac{\mu g}{mL}}{AU}\times\frac{100\mu L}{5\mu L}=158\frac{\mu g}{mL} }[/math]
[math]\displaystyle{ \frac{A_{260}}{A_{280}}=\frac{0.130AU+0.028AU}{0.055AU+0.029AU}=1.88 }[/math]
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