Alu PCR Lab- write up
This is the first lab we did this year. We took some DNA from our cheek cells and analyzed them in a gel, which showed us whether we had an alu segment in our DNA or not. It was a tiring process, but it was worthwhile with how much we learned.
PURPOSE
The purpose of this lab was to learn about the polymerase chain reaction. Polymerase Chain Reaction (or PCR) is used to amplify or make more copies of a single strand or couple strands of DNA. We used this method to amplify or make more of our cells to see if we had the alu segment in our DNA. We also learned about lab procedure.
HYPOTHESIS
My hypothesis for this lab was that not a lot of our class will have the alu gene.
MATERIALS
The materials we used for this lab are the following:
~ micropipettes (p20 and p200) ~ micropipette tips ~power supply
~ saline solution ~ microfuge tubes
~ our cheek cells ~ chelex
~ master mix (nucleotides,polymerase) ~ agarose gel
~ primer mix ~ DNA stain solution
~ micro centrifuge ~ heat block
PROCEDURE
We took the following steps to get our results:
PART 1
1) Swirl the saline solution in your mouth for 30 seconds
2) Spit the saline solution (now with your cheek cells) into a cup and stir the cup to mix the cells
3) Label a microfuge tube with your initials and put 1000 microliters of the saline and the cells in the tube
4) Spin the cell mixture in the micro centrifuge for 1 minute
5) There will be a cell pellet at the bottom of the tube. Pour off the extra liquid at the top and dispose it
6) Re mix the solution
7) Add 50 microliters of your cells into a tube of chelex
8) Place your chelex tube on the heat block and wait 10 minutes
9) After it's heated, release the pressure by opening up the tube, and spin it on the centrifuge for 1 minute
10) Take out 50 microliters of the top of the chelex tube and place it in a new tube
11) Refrigerate the new tube
PART 2
1) Place 20 microliters of the master mix in the new tube (mentioned above)
2) Place 20 microliters of the primer mix into the tube
3) Take 10 microliters of your DNA from your chelex tube and put it in the new tube
4) Heat up the new solution
PART 3
1) Measure 2 grams of agarose
2) Mix with a buffer solution until it is blended
3) Heat up the new solution
4) Pour the now hot solution in a box
5) Wait for the solution to set and cool
6) While you wait for it to cool, place a comb in the solution
PART 4
1) Place the new tube with your DNA in the centrifuge and spin it for 10 seconds
2) Add 5 microliters of loading dye in the tube with your DNA in it
3) Place up to 20 microliters (no less than 15 microliters) of your DNA and dye mixture in the gel
4) Add 5-10 microliters of a bp ladder (a molecular weight marker) to the end of the gel for balance
5) Attach the gel to a power supply for 40 minutes
6) Find the Alu gene
RESULTS
The result of my lab is +/-, meaning I had a portion of the alu gene from one of my parents.
This is the first lab we did this year. We took some DNA from our cheek cells and analyzed them in a gel, which showed us whether we had an alu segment in our DNA or not. It was a tiring process, but it was worthwhile with how much we learned.
PURPOSE
The purpose of this lab was to learn about the polymerase chain reaction. Polymerase Chain Reaction (or PCR) is used to amplify or make more copies of a single strand or couple strands of DNA. We used this method to amplify or make more of our cells to see if we had the alu segment in our DNA. We also learned about lab procedure.
HYPOTHESIS
My hypothesis for this lab was that not a lot of our class will have the alu gene.
MATERIALS
The materials we used for this lab are the following:
~ micropipettes (p20 and p200) ~ micropipette tips ~power supply
~ saline solution ~ microfuge tubes
~ our cheek cells ~ chelex
~ master mix (nucleotides,polymerase) ~ agarose gel
~ primer mix ~ DNA stain solution
~ micro centrifuge ~ heat block
PROCEDURE
We took the following steps to get our results:
PART 1
1) Swirl the saline solution in your mouth for 30 seconds
2) Spit the saline solution (now with your cheek cells) into a cup and stir the cup to mix the cells
3) Label a microfuge tube with your initials and put 1000 microliters of the saline and the cells in the tube
4) Spin the cell mixture in the micro centrifuge for 1 minute
5) There will be a cell pellet at the bottom of the tube. Pour off the extra liquid at the top and dispose it
6) Re mix the solution
7) Add 50 microliters of your cells into a tube of chelex
8) Place your chelex tube on the heat block and wait 10 minutes
9) After it's heated, release the pressure by opening up the tube, and spin it on the centrifuge for 1 minute
10) Take out 50 microliters of the top of the chelex tube and place it in a new tube
11) Refrigerate the new tube
PART 2
1) Place 20 microliters of the master mix in the new tube (mentioned above)
2) Place 20 microliters of the primer mix into the tube
3) Take 10 microliters of your DNA from your chelex tube and put it in the new tube
4) Heat up the new solution
PART 3
1) Measure 2 grams of agarose
2) Mix with a buffer solution until it is blended
3) Heat up the new solution
4) Pour the now hot solution in a box
5) Wait for the solution to set and cool
6) While you wait for it to cool, place a comb in the solution
PART 4
1) Place the new tube with your DNA in the centrifuge and spin it for 10 seconds
2) Add 5 microliters of loading dye in the tube with your DNA in it
3) Place up to 20 microliters (no less than 15 microliters) of your DNA and dye mixture in the gel
4) Add 5-10 microliters of a bp ladder (a molecular weight marker) to the end of the gel for balance
5) Attach the gel to a power supply for 40 minutes
6) Find the Alu gene
RESULTS
The result of my lab is +/-, meaning I had a portion of the alu gene from one of my parents.