DNA extraction.

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DNA extraction.

Postby anabellabonn » Wed Oct 17, 2012 9:36 am

I wanted to do a science project regarding DNA extraction.
Would it be possible to extract certain DNA to give another organism another trait, as in adding to their their DNA? Or if I could replace DNA sequences with other DNA sequences without risking any harm to any organisms?
I already have experience extracting DNA, and what laboratory safety hazards would this lab involve? What organisms would be fair to experiment on?

Recently, I have been thinking of trying to see if DNA extraction will be able to be used while detecting a certain disease or bacteria. I need more feedback and ideas.
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Re: DNA extraction.

Postby sunmoonstars » Fri Oct 19, 2012 8:03 am

anabellabonn wrote:I wanted to do a science project regarding DNA extraction.
Would it be possible to extract certain DNA to give another organism another trait, as in adding to their their DNA? Or if I could replace DNA sequences with other DNA sequences without risking any harm to any organisms?
I already have experience extracting DNA, and what laboratory safety hazards would this lab involve? What organisms would be fair to experiment on?

Recently, I have been thinking of trying to see if DNA extraction will be able to be used while detecting a certain disease or bacteria. I need more feedback and ideas.


Hi,

We have lots of DNA projects in the Science Fair Project Ideas - search using the keyword "DNA". I found this one that helps answer your questions: http://www.sciencebuddies.org/science-fair-projects/project_ideas/BioChem_p013.shtml#summary

Yes, it is possible to insert DNA into the cell's natural DNA. We usually don't replace DNA sequences with others, but we add new ones. We don't have much control over where the new DNA inserts into the natural DNA. A popular method used to insert DNA is called TOPO cloning and it is explained a little bit here: https://www.invitrogen.com/site/us/en/home/brands/Product-Brand/topo/The-Technology-Behind-TOPO-Cloning.html

Let me know what questions you have then.
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Re: DNA extraction.

Postby heatherL » Sat Oct 20, 2012 11:11 am

Hi anabellabonn,

As I mentioned in a reply to one of your other posts, you really need to keep all related posts in the same thread. This is your fourth separate post on the same topic, in two different forums!

Here is a link to my earlier reply, which should answer most of your questions. In it, I include a link to a Bio-Rad site that can help you perform the project you want to do!
viewtopic.php?f=28&t=10752&p=38259#p38259

Heather
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Re: DNA extraction.

Postby anabellabonn » Tue Oct 23, 2012 7:26 pm

I have decided to do a lab similar to this one regarding DNA purification and testing what enzymes are able to be used in the purification process. http://www.sciencebuddies.org/science-f ... ml#summary


What enzymes are preferably to be used in this experiment?
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Re: DNA extraction.

Postby heatherL » Wed Oct 24, 2012 6:48 am

Hi anabellabonn,

I'm glad you found something that interests you! The best thing for you to do is begin researching different DNA purification protocols.

I found this site when I put "DNA purification" in a Google search: http://www.protocol-online.org/prot/Mol ... index.html

If you use Google Scholar, you can find scientific articles that describe their DNA purification methods. Here is one I found by searching "DNA purification efficient" in Google Scholar: https://www.princeton.edu/genomics/bots ... _Rapid.pdf. There were many others!

Keep in mind that the project you've chosen is an extension of the onion DNA extraction project: http://www.sciencebuddies.org/science-f ... ml#summary. Your project talks about looking at different enzymes like protease (which degrades proteins), DNAse (which degrades DNA), and RNAse (which degrades RNA) to improve purification efficiency. If you are purifying DNA, you would want to avoid DNAse, since it will destroy the DNA. Using other enzymes (protease, RNAse) to break down the other macromolecules may help you.

Take a look at the different protocols and the different enzymes they use. You will probably want to focus on one or two enzymes (in addition to a control) that may increase purification efficiency, so you can replicate the process several times.

Let us know if you have more questions along the way!

Heather
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Re: DNA extraction.

Postby anabellabonn » Fri Nov 02, 2012 10:13 am

Regarding using other enzymes for the extraction, i haven't been able to actually find other enzymes related to Protease. Any suggestions?
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Re: DNA extraction.

Postby heatherL » Fri Nov 02, 2012 12:23 pm

I'm not sure exactly what you are asking here. Are you planning to try different proteases to see which one works best?

There are many, many different proteases out there.
Here is the Wikipedia article about proteases. http://en.wikipedia.org/wiki/Protease Make sure to use Wikipedia as a jumping-off point (refer to the Bibliography), since you cannot always trust the information in the article itself.

Here is a site from which you can order specific proteases. http://www.sigmaaldrich.com/life-scienc ... inder.html

Do you understand why a protease is used in DNA extraction? What do you think would make one protease work better than another? Will it depend on the organism from which you are extracting the DNA? Try answering these questions, which should help you in designing your experimental protocol.

Heather
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