Alu PCR Lab
Purpose: The purpose of this lab was to use micropipetting techniques to determine if you and/or your parents have the repeating gene sequence "ALU".
Hypothesis: I predict that my result will be +/-, or heterozygous. The result would mean that one of my parents has ALU (as indicated by the '+', and one does not (the '-'.)
Materials:
The materials used were:
Procedure:
Results: I am, as I predicted, heterozygous. One of my parents has the Alu sequence.
Analysis: My hypothesis was correct! The next step would be to test my parents and see which of them has Alu in their DNA.
Evidence of Completion:
Hypothesis: I predict that my result will be +/-, or heterozygous. The result would mean that one of my parents has ALU (as indicated by the '+', and one does not (the '-'.)
Materials:
The materials used were:
- 10mL Saline Solution
- Our own cheek cells
- A paper cup (for the saline solution and the cheek cells
- 1.5mL microcentrifuge
- Microcentrifuge
- Microcentrifuge tube rack
- Chelex tube
- Thermal cycler
- a P-20 and P-200 micropipette
- refrigerator
- 20mL Primer Mix
- 5mL loading dye
- Gel box
- Gel
- Gel comb
Procedure:
- Obtain cheek cells by using the saline solution to swish vigorously, then spitting the mixture of saline and cells back into the cup.
- Micropipette a small amount of the solution into a PCR tube, then spin the tube in a microcentrifuge to send the cheek cells to the bottom of the tube.
- Drain excess saline, to have only the cell pellet and a tiny amount of saline left.
- Add cell pellet to chelex.
- Apply cap lock and heat overnight.
- Take off the cap lock and place the tube back in the centrifuge.
- Transfer supernatant from that tube to a new one, and a refrigerate the new tube after.
- Spin the refrigerated tube in a centrifuge, put a small amount in a smaller PCR tube, and add the Master and Primer mixes.
- Place in the thermal cycler and wait overnight.
- Spin again in the micro-centrifuge, add loading dye, and put it in a hole in the gel. Send 150 volts of electricity through the gel fro 30 minutes, stain the gel, and photograph!
Results: I am, as I predicted, heterozygous. One of my parents has the Alu sequence.
Analysis: My hypothesis was correct! The next step would be to test my parents and see which of them has Alu in their DNA.
Evidence of Completion: