Alu PCR Lab:
Purpose:
-Successfully isolate DNA from cheek cells
-Prepare a PCR reaction for amplification of an Alu insert
-Prepare a PCR reaction for amplification of an Alu insert
Materials:
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Procedure:
1. We started by obtaining a sample of our own cheek cells to isolate DNA from. (using a saline wash)
2. We centrifuged our samples so the cells created a pellet at the bottom of our tube.
3. Before adding Chelex beads to our cell pellet we poured out the supernatant, whatever left in saline sample that was not cheek cells.
4. To burst the cell walls of our samples we heated the cells and Chelex mixture.
5. We were left with another pellet at the bottom of our tube and a clear liquid on top. The liquid was our isolated DNA.
6. We extracted our DNA and added it with master mix and primer mix into a tiny PCR tube.
7. All our samples were then loaded into a thermal cycler.
8. Gels were made for electrophoresis by combining 1xTAE and agarose until dissolved. We let it sit until it became a gel.
9. To prepare our DNA samples for loading into our gels we combined load dye and our DNA.
10. Marking which lanes were loaded with what, we loaded our samples into the wells or the gels.
2. We centrifuged our samples so the cells created a pellet at the bottom of our tube.
3. Before adding Chelex beads to our cell pellet we poured out the supernatant, whatever left in saline sample that was not cheek cells.
4. To burst the cell walls of our samples we heated the cells and Chelex mixture.
5. We were left with another pellet at the bottom of our tube and a clear liquid on top. The liquid was our isolated DNA.
6. We extracted our DNA and added it with master mix and primer mix into a tiny PCR tube.
7. All our samples were then loaded into a thermal cycler.
8. Gels were made for electrophoresis by combining 1xTAE and agarose until dissolved. We let it sit until it became a gel.
9. To prepare our DNA samples for loading into our gels we combined load dye and our DNA.
10. Marking which lanes were loaded with what, we loaded our samples into the wells or the gels.