Background: In todays modern world, we use Molecular biology, is the study of of genes and other molecular structures, that regulate the flow of genetic information from generation to generation. Also Biotechnology uses this information to help solve human problems, by taking this information and testing it through DNA tests, just like our very own lab. In our our lab we are doing a DNA test, to find out whether or not certian people in class have a specific gene that we are tryign to target. This DNA testing is used for all sorts of things, but it is most commonly used for helping people find out if they have or carry a certain disease like Huntington disease. In order to do this lab we must use PCR, which is a technique that takes a trace of DNA and produces a large amount of it. This technique today is widely used in biotechnology and it even helepd transfrom molecular biology into a multidisciplinary research field within five years of it being made. PCR has also had a profound impact on four main areas of genetic research: gene mapping, gene cloning, DNA sequencing and gene detection.
Directions:
Step 1. First we labeled one screw cap tube containing 200 hl of InstaGene matrix and then rinsed our mouth with saline solution for 30 secounds. (chewed inside of our gums to loosen cheek cells.).
Step 2. Then we placed this saline solution that we spit out into the cap tube containing the InstaGene and then we centrifuged it for about 2 minutes.(After it was centrifuged, there was a white bead of whitish cells at the bottom of the tube.)
Step 3. Then after centrifuging, we poured out the extra saline solution out of the tube, so then we only had the pellet of white cells at the bottom of the tube.
Step 4. Then we vortexed the tube so there would be no clumps of cells remaining and to resuspend the pellet of cells.
Step 5. Then we transfered the resuspended cells, into the screwcap tube containing InstaGene.
Step 6. Then we vortexed the tube and then placed the tube into a water bath (56 degrees Celcius) for 10 minutes.
Step 7. After we placed the tube in the vortex again and then put it into a boiling water bath (100 degrees Celcius) for 5 minutes.
Step 8. Then the next day we placed the mixture into a PCR tube and also added some yellow master mix. Then we placed it into the thermal cycler for 40 cycles.
Step 8. Finally, we added loading dye to our PCR tubes and then placed our mixtures into the wells of the agrose gel and then turned on the electrophoresis apparatus for about 10 minutes.
Results: Our results showed that we were all negative for the disease, but it could possibly be wrong because we could have messed up when loading the mixtures into the wells of the agrose gel. Since our results were very faint and very hard to see if they really were negative for the disease. I also think that my tube might have not had enough cheel cells because when I tried transfering the cheek cell bead to another tube, it sort of feel apart.So this may have caused me to transfer more of the saline solution rather than my cells.
