Telomerase assay.

[theroses]

For my science fair, I want to conduct and experiment to see the change in telomere length before and after a decided lifestyle: (diet, exercise and stress-level), to see what effect positive changes in lifestyle can have on telomere length. I am going to be doing this experiment on myself and a few other people.

To do this experiment, I was first considering conducting a PCR on telomeres folowed by a gel electrophoresis. This was my first option, but I do not have access to the specialized equipment necessary to conduct a precise enough PCR.
My next option is to conduct a telomerase assay, but, so far, all of the articles I have researched also include PCR, which is again not a viable option due to not precise enough equipment.

Is there a way to conduct a telomerase assay without the use of PCR?

[SciB]

Hi,

Telomere shortening is a hot topic for aging research now, so your project hits the mark for originality. The problem as you say is that measuring telomere shortening requires some pretty serious lab equipment, expensive reagents and knowledge of the methods involved. Also, you have a great idea in comparing the effects of different lifestyles on telomere length, but this is not something you can expect to see in a month or so—more like years I would think.

Human telomeres can be thousands of nucleotides long, but they don’t shorten that quickly, so the changes are small when looked at over a relatively short time period. You are trying to see changes in the length of telomeres in humans and that requires either PCR or DNA sequencing, both of which involve expensive equipment and reagents.

It is possible that you could isolate DNA from cheek cells of your volunteer friends, run it on an agarose gel, do a Southern blot and probe it with a sequence specific for telomeres. I’ll have to do some searching through PubMed to find out whether this is feasible for you to try. Making and running the gel is simple and does not require any elaborate or expensive equipment. In fact, gel boxes can be homemade. The difficult part is identifying the DNA band that corresponds to the telomere, and for that you would need to buy a labeled DNA sequence specific for the telomere. This could cost $50-100 depending on the length.

Let us know what you plan to do and we can try to guide you to a doable project with the resources you have access to. It is a wonderful project! If you could find a mentor and a lab doing telomere research, this would make a great study. Good luck!

Best wishes,

Sybee

[theroses]

Hello,
Thank you so much for your response. If I would have to buy DNA sequencing between 50-100 dollars, that wouldn't be a problem. Given that this experiment would take years as you said, do you know if there is any way to test the general health of a cell that would show more immediate results?

Thank you.

[deleted-71603]

Hi, theroses. I noticed you posted the same question in the Grade 6-8: Life, Earth, and Social Sciences forum. We deleted your post there, since you have received great advice on this post. Please keep all of your questions on the same topic on a single thread, as that will make it easier for us to help you.

Thanks for using Science Buddies, and good luck with your project!
Cheers,

[SciB]

Hi,

That’s a tough question to pin down because ‘health of a cell’ can apply to anything. I would really like to see you do something with the telomeres because that is such an interesting part of cellular aging. Since telomere shortening takes place over many years, you could compare telomeres from people your own age to those from people 20, 30 or 50 years older. Lifestyle, diet and environment as well as genetics play a part in how many years a person’s telomeres remain of sufficient length to protect the chromosomes.

Telomeres are attached to the ends of chromosomes, so to do a project to compare telomere length as a function of age and lifestyle you would need to isolate DNA from people ranging in age from 15 to 65 years of age or so. Do you have access to a high school or college/university lab where they do agarose gel electrophoresis of DNA? Can you ask someone to help you with the project as mentor? If so then I think you might be able to do a telomere comparison and relate it to the age and life style of your volunteer subjects. The one problem I don’t yet have a solution for is how to identify the telomere bands on a Southern blot. I think there must be some specific sequence in the telomere DNA that could be used as a probe, but this would be something you would have to research to find out. Hopefully one of the other experts can help with this part of the experiment. Here are some references I used:

What are telomeres?
http://learn.genetics.utah.edu/content/ ... telomeres/

Lifestyle changes may lengthen telomeres
http://www.ucsf.edu/news/2013/09/108886 ... cell-aging

Science Buddies project comparing human DNA from cheek cells to strawberry DNA
https://www.sciencebuddies.org/science- ... p042.shtml

Kit for isolating human DNA from cheek swabs
http://www.qiagen.com/products/catalog/ ... l-kit?pf=1

Agarose gel electrophoresis equipment
http://www.carolina.com/catalog/search- ... SearchForm



I’ve given you this idea but I’m not sure it will work. Of course, that’s the way a lot of science is done. If we knew everything ahead of time, where would be the fun in doing experiments? Ask your teachers and any friends you have at a college or university if they would be willing to help with your project. We can answer specific questions and provide you with sources of information, but you really need someone there with you to show you how to set up and run the gel, blot it and probe it for telomeres.

Let us know what you want to do and we will be glad to help in any way.

All the best,

Sybee

[deleted-137717]

Hey there,

I just wanted to chime in to show some support. This is an awesome project you're working on, and I think SciB's suggestion of comparing telomere length between age groups would be a great way to go about it. Definitely do try getting in touch with a nearby college or university. You never know what help and/or resources they might be willing to contribute. Keep us updated and good luck!

[deleted-132180]

Hello there,

Awesome project idea, and the experts have all given you excellent advice so far! I'd like to chime in with some of my thoughts.

I think it's very cool that you want to assess how diet, exercise, and stress-level affects the length of telomeres, but one of my concerns is that it may take a really, really long time to see the long term effects of these conditions on telomere length, like SciB had said. Furthermore, it would require you to find a large group of human volunteers who are on some specific diet, doing some specific exercise routine, and/or undergoing a similar stress, and this may be really difficult. I would suggest looking for a local lab who works on telomere length, talk with the lab members about the ideas you have, and you guys can brainstorm together to see how to make a project that is feasible for you to do. I think you have some very great ideas and you should definitely test them out, and looking for advice from experts on telomeres may give you a better sense of what you can accomplish.

Best of luck! Let us know if you have anymore questions.

Connie