You’re now ready to test your own DNA! Bring in your printed SNP Student Manual. As you know, this lab involves a number of steps. Below are the major steps, along with an estimate of how long they will take, and good stopping points.
Isolate DNA by saline mouthwash - takes approximately 45 minutes; can freeze samples or continue (continuing is suggested).
Amplify DNA by PCR - 20 minutes of hands-on work, then about 1.5 hour wait during thermal cycling. You can start your thermal cycling (PCR reaction) and ask staff to put it on the freezer for you when it’s done by leaving a note on the “Help, please!” board.
Digest PCR products with HaeIII - 15 minutes of hands-on work, then 30 minute wait while the restriction enzyme works. Can freeze after or continue.
Analyze PCR products by gel electrophoresis - approximately 2 hours, mostly waiting. Your end product will be a photograph of your gel.
Once you have a photograph of your gel, label it neatly so that your instructor or another student can easily tell what they are looking at. Note the labels for what’s in each lane, and for the sizes of the ladder band, that are highlighted in the image below. While you will only have three lanes, it’s important that they are all labeled neatly and clearly. In the assignment upload and submit your labeled figure.
Not the labels highlighted in red. You should similarly label your own gel photo.
Objective
Perform a single nucleotide polymorphism experiment and gel electrophoresis.