Abel Thomas

2/21/19

Purpose/Objective: The purpose and objective of this lab is to be able to see what type and how much DNA we have in our samples.

Materials:

Nanodrop

1X TAE buffer

31ng DNA mass

125ng DNA mass

250ng DNA mass

500ng DNA mass

10X leading buffer

Agarose Gel

Electrophoresis Gel

Nanodrop

DNA specimen

Procedure:

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
2. Select the walls in which the DNA will be put into
3. Extract 9 uL of DNA and transfer it into eppendorf tube
4. Add 1 uL of 10X loading buffer
5. mix solution
6. Add 10 uL of DNA from your DNA sample into the second well
7. Add 5 uL of DNA 31 ng mass to 5th well
8. Add 5 uL of DNA 125ng mass to the first well
9. Add 5 uL of DNA 500 ng mass to the third 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

Nanodrop:

1. Clean the machine and ensure the machine is clean and dry
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

Results:

Our DNA ended up being 722.3 ng/uL with 1.56/1.8 for the A260/A280 ratio and 1.14/2 for A260/A230 ratio. Our DNA peaked at 260 but with these numbers we can say we still had some proteins and carbohydrates in our sample which showed that our data was not pure.

Storage: We made sure to throw the gel away in the biohazard bag and made sure to rinse and dry everything carefully to ensure there was no contamination. After we did that, we were able to wipe down our lab area and made sure it was clean and good for the next group.

Conclusion/Future Goals: We were unable to get our sample to be pure DNA but that is okay because we were able to observe the techniques and protocol needed to observe the DNA we retrieved form our soil. Our future goal is to see how our results will end up compared to everyone else’s results. We will use PCR to identify and amplify the DNA we have and then later send it for it to be sequenced.