8-25-2015
ALU PV92 PCR LAB
OBJECTIVE
The first lab in a long list of labs in my sophomore year.
We discovered our alu gene and genotype in this experiment to see where our origins are from. These genes have relatively no meaning.
PROCEDURE 1ST DAY
1. Wash mouth in 0.9% saline solution for 30 seconds, then spit solution into cup
2. Pipette 1-1.5 mL. into microcentrifuge tube. Label tube and spin in centrifuge for 1 minute.
3. Pour out supernatant and resuspend cell bead at bottom of tube
4;. Pipette 50 uL into new tube with Chelex beads
5. Heat for 10 minutes.
6. Spin for 1 minute, add 50uL of supernatant to new tube
7) New tube- add 20uL of Master and Primer mix + 10uL of DNA. Put in thermal cycler.
PROCEDURE 2ND DAY
1) Mix 50 mL TAE with 1g agarose and heat until agarose dissolves
2) Wait for solution to cool and pour into mold. Remove comb when hardened.
3. Spin tubes, then add 20uL of DNA and 4uL of loading dye
4. Add DNA into hardened gel.
MATERIALS
0.9% saline solution
PCR tubes
1x TAE stock
Master Mix
Primer Mix
PCR tubes
Micropipette + tips
microcentrifuge + tubes
agarose
Gel chambers + molds
chelex solution
Water
Waste container
Tube racks
RESULTS
I am Negative Homozygous (Negative, Negative).
ANALYSIS
When we initially did this lab, less than half of the class wasn't able to get a correct analysis or answer on what type of alu genes they have. There were errors with transferring the solutions and telling detailed instructions. I was unable to read the gel clearly to get my result, so I had to restart the lab, and follow all of the steps again. This was frustrating but after we completed the lab again I was able to clearly see the result. The only bad part of the lab was swishing my mouth with the saline solution.
ALU PV92 PCR LAB
OBJECTIVE
The first lab in a long list of labs in my sophomore year.
We discovered our alu gene and genotype in this experiment to see where our origins are from. These genes have relatively no meaning.
PROCEDURE 1ST DAY
1. Wash mouth in 0.9% saline solution for 30 seconds, then spit solution into cup
2. Pipette 1-1.5 mL. into microcentrifuge tube. Label tube and spin in centrifuge for 1 minute.
3. Pour out supernatant and resuspend cell bead at bottom of tube
4;. Pipette 50 uL into new tube with Chelex beads
5. Heat for 10 minutes.
6. Spin for 1 minute, add 50uL of supernatant to new tube
7) New tube- add 20uL of Master and Primer mix + 10uL of DNA. Put in thermal cycler.
PROCEDURE 2ND DAY
1) Mix 50 mL TAE with 1g agarose and heat until agarose dissolves
2) Wait for solution to cool and pour into mold. Remove comb when hardened.
3. Spin tubes, then add 20uL of DNA and 4uL of loading dye
4. Add DNA into hardened gel.
MATERIALS
0.9% saline solution
PCR tubes
1x TAE stock
Master Mix
Primer Mix
PCR tubes
Micropipette + tips
microcentrifuge + tubes
agarose
Gel chambers + molds
chelex solution
Water
Waste container
Tube racks
RESULTS
I am Negative Homozygous (Negative, Negative).
ANALYSIS
When we initially did this lab, less than half of the class wasn't able to get a correct analysis or answer on what type of alu genes they have. There were errors with transferring the solutions and telling detailed instructions. I was unable to read the gel clearly to get my result, so I had to restart the lab, and follow all of the steps again. This was frustrating but after we completed the lab again I was able to clearly see the result. The only bad part of the lab was swishing my mouth with the saline solution.