User:Michaela Harper/Notebook/Biological Chemistry Lab/2011/09/21

From OpenWetWare
Jump to: navigation, search
BDLlogo notext lr.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png


To determine whether or not the synthesis of mutated GFP DNA was successful through gel electrophoresis.


  1. Add 0.25 g of 0.01 g/mL agarose to 25 mL of 1x tris base, acetic acid, and ethylenediaminetetraacetic acid (TAE) buffer. Heat the solution in a microwave for 40 seconds. The result is a 1% agarose gel used in electrophoresis. Pour the solution into the gel container with the appropriate dams. Allow the gel to cool for approximately 20 minutes.
  2. Mix 1 μL of 6x gel loading dye with 5 μL of the mutated DNA sample.
  3. Load the samples with dye mixture and a ladder containing DNA, glycerol, and gel loading dye into the wells of the gel. The ladder used was as follows: 500 bp, 1000 bp, 2000 bp, 3000 bp, 4000 bp, 5000 bp, 6000 bp, 7000 bp, 8000 bp, 9000 bp, 10000 bp.
  4. Run the gel electrophoresis at 80 V for 40 min.
  5. Remove gel and place in TAE and ethidium bromide (EtBr) solution, mix for 15 min.
  6. Rinse gel with TAE for 5 min.


All groups were successful in the synthesis of the mutated DNA. The desired product was 3600 bp. Using a UV light, we were able to see the DNA of all groups located between the 3000 bp and 4000 bp ladder spots, indicating a successful synthesis. The photo of the electrophoresis gel:

DNA gel 092111 003.jpg

The first column is the ladder. The next columns are groups 1, 2, 3, 4, and 5.