User:Brian P. Josey/Notebook/2009/11/25

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Digestion of pBR322

We are going through the process of preparing the pBR322 from yesterday. Yesterday, we were able to digest it with EarI. Today, we are going to gel extract it at the proper EarI site and then ligate it into three pieces. To this affect, I created a gel with 0.4g of agrose and 50 mL of the TAE buffer.

Gel Extraction

  1. Create a gel with 50 mL of TAE (from the bottle) and 0.4 g of Agarose. Use the larger comb
  2. Add dye to the DNA, one fifth of the volume, and mix.
  3. Add the DNA to the center well and add 30 μL of the ladder to the wells to the left
  4. Run the gel.