User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/05/26

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Today's Benchwork

  1. 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.
  2. Quantify [ pCMV-SPORT6+BSA ]
    1. 95μL H2O
    2. + 5μL miniprepped DNA from step 1
    • → measure absorbance at 260 nm and 280 nm
  3. PCR of BSA for cloning into pTXBI

Results

[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}

\frac{A_{260}}{A_{280}}=\frac{0.130AU+0.028AU}{0.055AU+0.029AU}=1.88


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