In this lab we began our identification of the murderer by first putting our tube with the restriction enzymes in an ice bath. Then we collected our evidence from the crime scene, in five test tubes. We had narrowed it down to five suspects! We got a sample of each suspects DNA and placed them in test tubes, then we added 10 ul of the enzyme mix into the samples and mixed well. Next, we placed the test tubes into an incubator at 37 degrees Celsius for 45 minutes. Mr. Chugh took out our test tubes at the correct time and got them ready for when we returned the next day.
The next day when we came back we removed the tubes from the water bath and then we gentle tapped the tubes on the table to bring the liquid to the bottom of the tubes. Next, we added 5 ul of loading dye into the separate tubes, collecting the samples in the end of each test tube. We took the agarose gel and placed it in the electrophoresis apparatus. We checked that the wells of the agarose gels were near the black negative electrode and the bottom was near the red... But we found that our positive and negative was switched! So, we replugged the electrophoresis to correct the error. The next thing we did was using a separate tip for each sample we loaded the DNA samples into the gel and after putting the lid over the chambers we turned on the power. When we returned the next day, we could finally compare our results! We found the real murderer and solved our simulated crime scene investigation.
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