Sydney Ortenberg

Feb 22, 2019

Lab 6: Running the Gel

Objective/Purpose:

The objective of this lab was to run the gel electrophoresis and analyze our DNA within the gel. The purpose of this lab was to hopefully find DNA within our sample. The nanodrop technology would analyze our sample and see how pure it is and if there was any contamination.

Procedures:

Gel Electrophoresis

1. Remove the gel from the mold and transfer it onto the transfer dock and cover the gel and the rest of the docker with 1X TAE buffer (with gloves on)
2. Select the walls in which the DNA, mass standards and practice buffer will be put in.
3. Extract 9 uL of DNA and transfer it into Eppendorf tube
4. Add 1 uL of 10X loading buffer to the Eppendorf tube
5. Mix solution with the vortex for 1o seconds.
6. Add 10 uL of DNA from your DNA sample into a well
7. Add 5 uL of DNA 31 ng mass to a well
8. Add 5 uL of DNA 500 ng mass to a well
9. Add 10 uL of DNA from the other group’s sample to different well
10. Run the agarose gel at 100 V for 20 minutes
11. Take the gel to the UV illuminator and observe the image to see how much DNA was found in your sample by comparing it to the mass standards.

Nanodrop Technology:

1. Clean the machine and ensure the machine is clean and dry. Neutralize the machine by placing a drop of water on the machine first, and running it.
2. Place 1 uL of DNA sample on the top of the holder
3. Close the machine and wait for results
4. Analyze the results that appear on the screen.

Data and Results:

• we do not have access to the images of the Gel under the UV light

Storage:

The gel mold was disposed of and all other equipment was stored away. The gel mold container was bleached and cleaned and put in the correct area.

Conclusion and Future Goals:

In conclusion, we were able to detect DNA in our sample from gel electrophoresis. The agarose gel showed us the different sizes of our DNA bands compared to our mass standards, and we were able to view them. More analysis will take place that will allow us to identify our band sizes more specifically. We were also able to use a Nanodrop spectrophotometer to gather data relating to the concentration and purity of DNA present in our sample. Using this, we found that our DNA contained some impurities. DNA should be at 1.8 and our sample was 1.61. In the future, we will be figuring our DNA bands more exactly, and comparing them to others, and we will also be sending them off to be analyzed and grouped together through bioinformatics.