Overview
For this lab, I worked with Tony, Naomi, and Pokie. Our goal was to test our DNA, and find out if we had the 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 using the PCR in our DNA, then we can learn who is heterozygous or homozygous.
Materials:
Cheek cells
10mL of saline solution
Microfuge tubes
Microcentrifuge
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. Microcentrifuge 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. Observe results
Results
My results came out as -/- (homozygous), which means I do not have the alu gene. Here is a picture of my results. I am the very bottom strand:
For this lab, I worked with Tony, Naomi, and Pokie. Our goal was to test our DNA, and find out if we had the 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 using the PCR in our DNA, then we can learn who is heterozygous or homozygous.
Materials:
Cheek cells
10mL of saline solution
Microfuge tubes
Microcentrifuge
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. Microcentrifuge 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. Observe results
Results
My results came out as -/- (homozygous), which means I do not have the alu gene. Here is a picture of my results. I am the very bottom strand: