Team:CSU Fort Collins/Notebook/Protocols=Gel

From 2014.igem.org

CSU iGEM 2014

Gel Electrophoresis Protocol

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  1. Using a balance, mass out 1.0 gram of agarose. Mix this with 100 mL of TAE 1X Buffer.
  2. Microwave the mixture, mixing intermittently, until all the agarose is dissolved and the mixture is homogeneous.
  3. Pour the mixture into a gel plate and insert an 8-lane comb. Allow to cool and solidify in the fridge or on the benchtop
  4. Mix your samples to run in the gel. Use 10 μL of the DNA Ladder combined with 2 μL SYBR Green in the first lane and for all samples, mix 5 μL of sample with 5 μL of nuclease-free water and 2 μL of SYBR Green/Dye Mixture.
  5. Place gel in tray into the gel electrophoresis apparatus; Wells should be located towards the negative (conventionally black) end. Fill apparatus with 1X TAE until the gel is completely submerged. Do not overfill.
  6. Load 10 μL of each sample into the appropriate wells.
  7. Connect wiring and run gel electrophoresis for 1 hour.
  8. Analyze gel in UV light and see if samples match the expected size using the ladder as a guide.



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