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, and/or 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. They include image credits as well as research citations.
A free version of Camtasia was used. We apologize for the watermark.
Citations for this video (information and photo credits) are accessible in this document: biotech_citations_aldhia .
Excellent work. Good experimental design and concise presentation of the work. My question is that how do you validate your results? If you don`t observe a mutation, is it due to primer design or not optimal PCR conditions rather than a lack of mutation?
The answer to your first question: If SNP occurs in a non-coding region, it will not be translated into the protein. Even SNP is in a coding area, it may not affect the activity of the enzyme if the mutation is not in the catalytic site of the enzyme.
Dr. Bermek,
Thank you so much for watching our video and looking at our research! The region we amplified include one of the three SNPs in ADH1A; the amplified region could be cut in two with a restriction enzyme if the SNP was present in the subject’s DNA. Or, if the materials were available, the amplified region could be sequenced to determine if the SNP were present. Alternatively, new primers could be developed where the SNP was within the primer and without the SNP there would be no amplification at all.
Outstanding video. Very good: experimental plan, primer design, explanation of how you will check your amplicon, explanation of different genes in the gene family, and pronunciation of challenging vocabulary.
Video Production: Nice overall. One point- the video stays on the Chromosome 4 image for quite some time and then quickly switches to another image and then back to the Chromosome 4 image. This transition is awkward. Next steps: It is now time to add your final gel images with a results section describing PCR (Master Mix), PCR cycle, and electrophoresis. Great: Audio, Use of images for explanation as a whole, and initial images of the two of you!
Science: 1.) Although you are amplifying the ALDH1A gene and can check the amplicon using a restriction enzyme digest- how would you test for one of the SNPs in the area? What would be your experimental design for this? 2.) Are there ALDH genes in other areas of the genome? Is there any work that indicates which genes (multiple) are more positively correlated with alcoholism? How would you conduct a larger study to determine this?
Dr. Todd,
Thank you for all of your help on this project. We could not have done it without you! In order to determine if there is an SNP in the amplified region there are three options. The first if to find a restriction enzyme which cuts at the SNP, which would only cut in the presence of the SNP. The second is to have the amplified region sequenced. The third is to develop a new set of primers which would incorporate the SNP into the primer, so that any amplicon would show there was or wasn’t an SNP in the ADH1A gene.
Overall, there are a very large number of genes associated with alcoholism and addiction in general. But, in the chemical reactions which converts ethanol to acetaldehyde and acetaldehyde to acetate in the body there are two types of chemicals: alcohol dehydrogenases and acetaldehyde dehydrogenase. All alcohol dehydrogenase genes (ADH1A, ADH1B1, ADH1B2, ADH1B3, ADH1C1, ADH1C2, ADH4, ADH5, ADH7) are on chromosome four within 370 kbp. Mutations in these genes vary, with some increasing the risk for alcoholism whilst other mutations prevent alcoholism. Three Acetaldehyde dehydrogenase genes (ALDH1A1, ALDH1B1, ALDH2*1, ALDH2*2) are most related to alcoholism, but there are 18 total ALDH genes. These genes, unlike the ADH genes, are scattered throughout the genome. Because of the sheer quantity of genes related to alcoholism, breaking experiments down into multiple individual gene based experiments is the best way to determine which genes are most important in addiction.
Video production- Well done! Good intro and discussion regarding why study was important. Infographic was a little hard to read- blurry. Consider highlighting areas on slide/graph that you want to draw the learner’s attention to.
Thank you, Tina, for taking your time to watch our video and review our research! As we make more videos, we will make an effort to use higher quality images and highlight important information.
Your work is exceptional and way beyond that most high school students. It is easily beyond the caliber of undergrad students. Keep up the fine work biotech. The integration of computer skills and research work/presentation will last you a lifetime.
Excellence is what we strive for here at Cary Academy. Thank you for taking the time to watch our video, we hope you found it interesting and enjoyable.