Gel Loading Lab
Each student will prepare the following samples to load into three different wells of an agarose gel:
Tube 1
5 ul dH2O
1 ul 6X loading dye
Tube 2
4 ul dH2O
1 ul 6X loading dye
1 ul DNA ladder
Tube 3
4 ul dH2O
1 ul U-View (6X loading dye +
DNA stain)
1 ul DNA ladder
After adding the last reagent to each tube, pipet the mixture in each tube up and down several times to
mix. (You can mix the reagents using the same tip used to add the last reagent to that tube.) If reagents
adhere to the sides of the tube, use a centrifuge to pool the mixture at the bottom of the tube.
Place your gel tray in the gel tank with the wells on the black (-) side. (Remember that DNA is negatively
charged so the DNA samples move toward the red (+) electrode.)
Use your stock solution of 10X TAE to make enough 1X TAE buffer to cover your gel.
Pour 1 X TAE buffer into the gel tank until the buffer just covers the gel. Once the gel is covered in
buffer you can pull out the comb.
Load 6 ul of each sample into a separate well.
Run the gel at 130 V for 30 minutes.
In your lab notebook be sure to record which sample was loaded into which lane so that you can identify
which samples you loaded and which samples your lab partner loaded onto the gel.
After electrophoresis is finished remove gel from tank, but leave gel on tray to transport gel to the
transilluminator. View gel with UV light and print a picture. Include the picture in the data section of
your lab write-up.
You can empty the used buffer into the sink. Rinse gel tank, gel tray, and comb with tap water and dry
with a paper towel. Clean up your lab station.