restriction enzymes and recombinant DNA

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i need help!!!
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restriction enzymes and recombinant DNA

Post by i need help!!! » Sat Dec 04, 2004 2:40 pm

I had to do a project for biology in which i had to model the process of using restriction enzymes and plasmids to form recombinant DNA in an insulin gene. I am having trouble with these two questions though. Can anyone help me??

1. Why would you want your restriction enzyme to cut the plasmid in only one location?

2. Why would you want your restriction enzyme to cut as closely to the insulin gene as possible without cutting into it?

If anyone can help me with this it would be greatly appreciated.

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biostudent84
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Post by biostudent84 » Sat Dec 04, 2004 5:42 pm

It all has to do with conservation of DNA. If you cut only once, and as close to the insulin gene as possible, you are able to keep take out the insulin gene and very little else. Every cut hat the potential to lose DNA due to waste.

How do we survive even though we can potentially lose DNA? Humans only use 10% of our DNA. The rest has actually been termed "Junk DNA." Some flowering plants can use as little as 1%.

i need help!!!
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Post by i need help!!! » Sat Dec 04, 2004 7:08 pm

thank you

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Post by biostudent84 » Sat Dec 04, 2004 10:51 pm

Cheers=)

Kyle

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Re: restriction enzymes and recombinant DNA

Post by shokoohi » Sat May 07, 2005 10:59 am

i need help!!! wrote:I had to do a project for biology in which i had to model the process of using restriction enzymes and plasmids to form recombinant DNA in an insulin gene. I am having trouble with these two questions though. Can anyone help me??

1. Why would you want your restriction enzyme to cut the plasmid in only one location?

2. Why would you want your restriction enzyme to cut as closely to the insulin gene as possible without cutting into it?

If anyone can help me with this it would be greatly appreciated.

hi I am farhad,a molecular genetic student,
about first question, the celected enzyme must cut plasmid just in one side becuse when you phorese digestion product you can see one band, but if you used two enzyme you will have tow band and when you want to ligate the insertion DNA it would be missmached between the plasmid part that could religate with plasmid, so if you have to digest your DNA insertion with two enzyme you have to used other methods.

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