objective/hypothesis
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materials
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In this lab, we used the method of PCR or Polymerase Chain Reaction to see if we had the alu insert or not. The purpose of this lab was to determine the frequency for our alu insert or in other words, if it was heterozygous or homozygous.
procedure
1. Get DNA by swishing saline solution in your mouth and spitting it into a cup.
2. Transfer 1000 uL into a labeled tube 3. Spin solution until small pellets from 4. Pour out supernatant, but don't lose pellet cell 5. Rack or flick tube to mix cells 6. We took 50 microliters of chelex and put it into our mix 7.Put the tubes in a heat block for 10 minutes 8. Extract50 microliters of this and separated this 9.Put 20 microliters each of the master and primer mixes into the separated bit 10. Place tube in thermal cycle 11. Spin tube for ten seconds 12. Add dye to PCR tube 13. Make gel and let it harden 14. Load DNA mixture into gel 15. Put gel in gel box 16. Turn power on for 20-45 minutes in 150 volts 17. Stain gel and photograph ConclusionWhile learning about our own DNA was very intriguing, I felt that the most important part of this lab was learning about the basic procedures in a biotech lab. For example, my alu insert was indistinguishable the first time around so I had to restart the entire process to get a viable result. This is similar to when scientists are taking part in an experiment, that they have to be persistent and learn from your mistakes to gain greater knowledge. The lab also taught me a great deal about pipetting, microfuge tubes, centrifuges, and other standard laboratory tools.
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Pipet
0.9% saline solution Student cheek cells 5% chelex Primer mix Master mix TBE buffer Agarose gel, tracking dye DNA stain Positive control DNA Base pair ladder results
On my first attempt at trying to find if I was heterozygous or homozygous, my results were inconclusive. On our gel, my DNA path never showed thus making me have to redo the whole procedure all over again. In the image below, it should have showed in lane three, but it did not.
In my second time through, I received the result of double negative homozygeous as seen in lane 2 in the image below.
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