In this lab, my table group was Leo, Angus, Anthony, Ryan, and Aidan. Our project was testing our DNA, and finding out if we had a specific Alu gene.
Purpose: The purpose of this lab was to separate DNA from a double strand to a single strand. This is done by using PCR, standing for Polymerase Chain Reaction. With DNA separated, it can be determined whether an individual has an Alu insert, and if they are homozygous or heterozygous.
Hypothesis: If we can follow the procedure correctly and succeed in using PCR on our DNA, then we can learn which of the people in our class are heterozygous and homozygous.
Materials:
cheek cells
10mL of saline solution
microfuge tubes
microcentrifuge
a rack
chelex
heat block
pipet
20uL of master mix (nucleotides, DNA polymerase)
20uLof primer mix
thermal cycler
5uL of loading dye
agarose gel
5-10uL of bp ladder
gel box
Procedure:
1. swirl 10mL of saline solution to get our cheek cells
2. label microfuge tube with initials and transfer 1000uL of the saline solution into it
3. spin your sample in a microcentrofuge
4. pour off the supernatant from the top of the microfuge tube and rack the cell pellet
5. obtain a tube of chelex and add 50uL to the chelex
6. heat the mixture in 99*C for 10 minutes
7. centrifuge the heated mixture for 1 minute
8. take out 50uL from the chelex/DNA without any beads and refrigerate the DNA
9. put 20uL of master mix and 20uL of primer mix together with 10uL of your extracted DNA
10. place the mixture in a thermal cycler
11. create agarose gel
12. microfuge the DNA and add 5uL of loading dye to the PCR tube
13. load 15-20uL of the DNA to the agarose gel
14. place it in the gel box and electrophorese the sample at 25-40 volts
15. photograph and observe results
Results:
The main result that I got was that I am negative negative. This means that I do not have the Alu insert. About half of the class was homozygous, like me, without an Alu insert. The rest of the class is heterozygous, which means they do have the Alu insert. It's very interesting to me how it's something you can test so easily in a classroom, and yet, it seems that you would need to be in a serious lab to test someone's DNA.
Analysis:
Not everybody has an Alu insert. The actual insert does not change anything serious in a human, but it is there for some people. In this lab, I learned how to follow instructions in a packet, and I became comfortable with using a micropipet. I felt like I was still a little questionable with how completely accurate my measurements were, as my results were vague and hard to tell completely.
Purpose: The purpose of this lab was to separate DNA from a double strand to a single strand. This is done by using PCR, standing for Polymerase Chain Reaction. With DNA separated, it can be determined whether an individual has an Alu insert, and if they are homozygous or heterozygous.
Hypothesis: If we can follow the procedure correctly and succeed in using PCR on our DNA, then we can learn which of the people in our class are heterozygous and homozygous.
Materials:
cheek cells
10mL of saline solution
microfuge tubes
microcentrifuge
a rack
chelex
heat block
pipet
20uL of master mix (nucleotides, DNA polymerase)
20uLof primer mix
thermal cycler
5uL of loading dye
agarose gel
5-10uL of bp ladder
gel box
Procedure:
1. swirl 10mL of saline solution to get our cheek cells
2. label microfuge tube with initials and transfer 1000uL of the saline solution into it
3. spin your sample in a microcentrofuge
4. pour off the supernatant from the top of the microfuge tube and rack the cell pellet
5. obtain a tube of chelex and add 50uL to the chelex
6. heat the mixture in 99*C for 10 minutes
7. centrifuge the heated mixture for 1 minute
8. take out 50uL from the chelex/DNA without any beads and refrigerate the DNA
9. put 20uL of master mix and 20uL of primer mix together with 10uL of your extracted DNA
10. place the mixture in a thermal cycler
11. create agarose gel
12. microfuge the DNA and add 5uL of loading dye to the PCR tube
13. load 15-20uL of the DNA to the agarose gel
14. place it in the gel box and electrophorese the sample at 25-40 volts
15. photograph and observe results
Results:
The main result that I got was that I am negative negative. This means that I do not have the Alu insert. About half of the class was homozygous, like me, without an Alu insert. The rest of the class is heterozygous, which means they do have the Alu insert. It's very interesting to me how it's something you can test so easily in a classroom, and yet, it seems that you would need to be in a serious lab to test someone's DNA.
Analysis:
Not everybody has an Alu insert. The actual insert does not change anything serious in a human, but it is there for some people. In this lab, I learned how to follow instructions in a packet, and I became comfortable with using a micropipet. I felt like I was still a little questionable with how completely accurate my measurements were, as my results were vague and hard to tell completely.