Purpose:

The purpose of the lab was to perform gel electrophoresis of the DNA sample, gel and positive and negative controls that was performed last week, also we peer reviewed our group’s introduction to make changes into making it better.

Procedure:

Gel Electrophoresis:

1. Prepare 300ml of 1X buffer solution using 10 TAE stock solution and water (30ml of 10X TAE stock solution has to be mixed with 270ml of water).
2. Measure 270ml of water using a measuring cylinder and transfer the water into a conical flask. Then, transfer 30ml of 10X TAE stock solution into the same conical flask using a serological pipette.
3. Prepare 30µl of 1X loading dye using 6X loading dye and water (5µl of 6X loading dye and 25µl water).
4. Carefully place the agarose gel into the gel box, ensuring that the gel is placed in such a way that the wells are nearer to the negative electrode.
5. Connect the wires from the gel box to the power supply, ensuring that the same colored cord is plugged into the same colored plug.
6. Carefully pour the buffer solution from step 2 onto the gel, so that the buffer solution covers the gel completely.
7. Practice loading a well with 5µl of loading dye from step 3.
8. Using a p10 micropipette, load a well with 5µl of the DNA ladder.
9. Using a p10 micropipette, transfer 10µl of soil DNA sample into a microcentrifuge tube. Micropipette 5µl of ethidium bromide into the same tube.
10. Repeat step 9 using the positive control and the negative control in place of soil DNA sample.
11. Using a p10 micropipette, load a well with 10µl of soil DNA sample.
12. Using a p10 micropipette, load a well with 10µl of the positive control solution.
13. Using a p10 micropipette, load a well with 10µl of the negative control solution.
14. Secure the lid on top of the gel box and switch on the power.
15. Adjust the settings on the power supply to run for 30 minutes at a voltage of 110V.
16. Remove the gel after 30 mins for analysis of the gel under UV light, so as to examine the bands of DNA present in the sample DNA.

Peer Review:

1. Exchange your introduction draft with a group member.
2. Read and write suggestions that could be used to better the draft.

Data:

• Wells 1-3 contained trials (one of each member of my group)
• Well 4 was unloaded
• Well 5 contained the negative control
• Well 6 contained the DNA sample
• Well 7 contained the positive control
• Well 8 contained the DNA ladder

The photo above is the UV imaging of my group’s gel.

The photo above is how the gel looked after gel electrophoresis was performed.

Conclusion:

In conclusion, my group was not able to observe any bands for the DNA sample which means there was an error at some point of the experiment in past labs. My group will try to figure out our problem in a future lab to see where we went wrong and if there is anything to do in order to fix it.