User:Nader A. Khamis/Notebook/Experimental Biological Chemistry AU Fall 2011/2011/09/21

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Objective

  • 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.

Description

  • To produce the gel, the following procedure was made.
  1. 0.25g of Agarose (0.01g/ml).
  2. Agarose was mixed with 25ml of 1x TAE buffer.
  3. The mixture was placed into a microwave for 40 seconds.
  4. 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.

Data

  • 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.

Image: DNA_gel_picture_-_Septeember_2011.jpg

Notes

This area is for any observations or conclusions that you would like to note.


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