Detection of Bt gene in Corn Through PCR - High School Science Fair - (Oct/25/2005 )

Hi my name is Doug and I am a student at North Springs High School in Atlanta, GA and I am currently enrolled in an independent studies class directed towards Science Fair. My experiment is to use PCR and electrophoresis to determine if the gene from Bacillus thuringiensis that is used in Bt Corn is present in the Corn's DNA. I have found procedures to extract the DNA from the corn but not to purify it for use in PCR. Also, I have not been able to find a Primer that codes for the Bt gene. Do I need to purify the DNA in order to use it in PCR? If I do I would appreciate it if someone could provide me with a procedure to do that. Lastly, If anyone knows where I could purchase the above mentioned primer that would also be appreciated. Thanks for your help.

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first of all, awesome science fair project!


? what is your DNA extraction protocol? those usually give you DNA pure enough to use in PCR

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try this link:

http://www.okccc.edu/bbdiscovery/documents...CRGM%20Food.htm

that may be what you seek, if your BT gene had the same source

if that doesnt' work, try a search for "primers PCR bt" on www.google.com and a ton of stuff pops up

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Nice project!

I will get back to you tomorrow when I have access to online journals. I should be able to dig up the sequence for the primers (BT-R1 cadherin) then.

-Matt

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First of all thanks for the replies! I've been searching for things on google for weeks and haven't gotten the results I did using those words... I guess I'm not very good at google. The link that was provided did not mention where the primers were purchased and the DNA extraction methods mentioned used higher powered centrifuges than I have access to. I found a product made by Sigma that looks like it would work, here is a link:

http://www.sigmaaldrich.com/catalog/search...ail/SIGMA/XNAP2

If anyone has any experience with this product or knows anything about it please let me know. Also, if anyone knows a good source to buy the primers from that'd be great too.

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Hey Doug! Your project sounds really great! I would love to help, but unfortunately, I know nothing of this subject. Hopefully you'll get that primer sequence!

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Hi Doug,

I've used this Extract-N-Amp kit from Sigma, and it's a piece of cake. Extraction takes minutes and you can use the extract directly in your PCR reactions. I have had a few issues with PCR inhibitors present in my extract (I'm doing mouse tail clips), but this problem was solved by simply diluting the extract 1:3 with a 50:50 mix of the extraction and neutralization buffers.

As far as the primers are concerned, there are several good companies that make PCR primers (IDT, MWG, Proligo). Here's a link to a GM Crop testing protocol in which they give the sequence of forward and reverse primers to the Cry1A gene of Bacillus thuringiensis.

Best of luck to you... very cool project!
Alicia

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