Procedure1. Swirl 10 ml of saline solution in your mouth for 30 seconds. Spit out saline into a cup and swirl to mix cells.
2. Transfer 1000 microliters of the saline/cell suspension into your labeled microfuge tube. Spin in a microcentrifuge to pellet the cells. Pour off the supernatent, allowing 100 microliters to cover the cell pallet. Rack the sample. 3. Withdraw 50 microliters of your cell suspension and add it to a tube containing Chelex. 4. Apply Chelex tube to a heat block for 10 minutes. 5. Remove Chelex tube from heat block. Use a P-200 to withdraw 50 microliters of supernatent from the Chelex tube and transfer to a fresh tube. 6. Obtain a tiny PCR tube and keep on ice. 7. Pipette 20 microliters of Master Mix into the PCR tube. Then add 20 microliters of Primer Mix. 8. Add 10 microliters of your extracted DNa into the PCR tube. 9. Place reaction into a thermal cycler. 10. Retrieve PCR tube and spin in a microcentrifuge. Then, add 5 microliters of loading dye. 11. Create and pour gels. Add 1XTAE solution. 12. Load 15 to 20 microliters of the DNA/loading dye mixture into a well in your gel. 13. Load 5 to 10 microliters of a 100 base-pair ladder (molecular weight marker) into the one well in each gel for later comparison. 14. Run gels. |
ResultsThe results show that I have one parent with the gene and one without the gene. So I am a +/-
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AnalysisThis was a fun project because I got to learn how to use several new types of scientific machinery.
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