Purpose: To write a rough draft of the introduction to our protocol article and to review the methods for running an electrophoresis gel

Materials:

• Ladder solution
• Gel electrophoresis chamber
• PCR product
• Positive/negative control
• UV imager
• TAE
• D.I. water
• Power supply
• DNA loading buffer (dye)
• Centrifuge tubes
• Micropipette

Procedure:

1. Add 30 mL of 10X TAE to 270 mL DI water in order to make the buffer
2. Take the gel created last week and place it in between the positive and negative yield in the gel electrophoresis chamber. Pour buffer over the gel and into the chamber.
3. Add 5 ul of ladder into a well using a micropipette. This will be the comparison sample for the controls and the experiment sample.
4. The PCR products will be the positive and negative control as well as the experiment sample. These should be in 3 separate centrifuge tubes to which 5 ul of dye will be added.
5. Take 10 ul of each PCR product and insert into 3 separate wells in the gel using a micropipette.
6. Place the lid on the chamber with the positive and negative electrical wires connected. Set power supply to 110 volts for 30 minutes.
7. Carefully remove the gel from the chamber and place on a UV imager to observe DNA strands.

Conclusion/Results:

Unfortunately, there were not any PCR products visible in our gels, even for the positive control, therefore we obtained negative results. When we meet next, we will need to decide what went wrong in our protocol and what needs to be adjusted. Perhaps this means discovering other ways of amplifying ciliate DNA and thus researching their diversity in soil. Research on obtaining a good DNA sample from ciliates and methods of sequencing this obtained DNA needs to be researched more thoroughly.