- Perform Gel Electrophoresis on PCR products prepared on 9/20/2011 to determine whether or not GFP was successfully mutated to contain a cystein residue after the enterkinase cleavage site.
- To produce the gel, the following procedure was made.
- 0.25g of Agarose (0.01g/ml).
- Agarose was mixed with 25ml of 1x TAE buffer.
- The mixture was placed into a microwave for 40 seconds.
- The hot mixture was poured into tray to form gel sheet and was left to cool.
- Approximately 25ul of pink wax was removed from DNA sample.
- 1ul of 6x loading dye was placed on parafilm.
- The loading dye was mixed with 5ul of DNA.
- All of the DNA samples were placed in gel from wells 1 to 6.
- The Ladder was in well 1.
- The gel was run at 80V for 40 minutes.
- The gel was carefully removed and placed in ethidium bromide and TAE buffer solution.
- The gel was washed with TAE buffer for five minutes.
- Observation of the gel indicated the presence of faint bands at the right side of the 2kb band in the ladder.
- From top to bottom, the band sizes for the ladder are 10, 8, 6, 5, 4, 3, 2, 1.5, 1, and 0.5 kilobases.
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