This is the first independent project in Cary Academy ADV Biotechnology. Consider watching this video and commenting to provide feedback. Please focus on the science, information-explanation, and/or the video production. At the end of the video, the students ask questions to the viewers. Citations can be found on a separate document underneath the video or within the video.
You may want to use headphones to hear this video (the audio is soft- additionally a free version of Camtasia was used.) We apologize for the watermark.
Works cited: works-cited-huntingtons .
Outstanding video. Very good: experimental plan, current research, existing testing, etc.
Video Production: Next steps- It is now time to add your final gel images with a results section describing PCR (Master Mix), PCR cycle, and electrophoresis. You could add the challenges that you encountered. Great: Excellent explanation and it is clear that you understand the experimental design. The image of the two of you at the beginning is awesome.
Science: 1.) How is it that you will confirm that you got the correct amplicon other than the size? 2.) In the image of the primer location on the sequence, something is wrong. You seem to have tried to indicate where the primers bind on the sense (5′-3′) strand which can not be accomplished unless you show the reverse compliment. 3.) Great image of the electrophoresis results but I do not understand how if there was a gain of repeats- that the amplicon got smaller? Is this an error in the image? 4.) Did you consider the challenges of amplifying a high GC repeat area before your primer design? What would you do differently? What does the research suggest in terms of diagnostic assay?
Thank you, Dr. Todd! Here are the answers to your questions respectively!
1. We could extract the DNA from the gel and conduct a DNA isolation.
2. Something indeed was wrong! We changed primers, but we believe due to the high GC concentration, we were still unable to amplify the region.
3. That was an error that we overlooked. With more base pairs, it should be further up on the gel.
4. We considered the challenges before we proceeded with the experiment, but we still wanted to do our best to try to amplify the section. For diagnostic testing, labs use a FISH test to diagnose Huntington’s disease; something we are unable to access and use.
Excellent work!
When I watched the video, I was a bit confused by your primer design. The reverse primer is not complementary to the sense DNA. I couldn`t find a region on the sequence you show where your primer could attach. Could you show in this sequence where your reverse primer attaches?
Video production- Excellent!! Your choice of graphics was very good and you highlighted areas of interest or focus for the viewer- particularly one who does not have an extensive science background. Pictures made it personal and your questions at the end reinforced this. Great job illustrating design and instrumentation of the project. In the future, be sure to check volume levels on recording for more even transitions between speakers.
The fact that you pointed out your lack of success with the outcome of the experiment is very significant. Few if any experimental research ever works out initially. This is one of the single most important scientific values to be learned. Persistence and repetition, review of procedures, and unbiased effort is very important. Confirmation of procedures and results by another team to see if data can be replicated is very important. Your work is outstanding and beyond that of most undergrad students. Please keep up the great effort of advanced research and computer skills to communicate your work.
Bill, thank you so much for your comments. My partner and I were very humbled by this experiment. Dr. Todd certainly helped us through our failures but your suggestion of having another group run the same test would have been a good idea and would have given us someone to compare our findings too. Again, thank you so much for your response to our video.