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help in digesting DNA fragment using restriction enzyme

restriction enzyme DNA digestion ligation

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2 replies to this topic

#1 minerva_29

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Posted 07 October 2011 - 09:20 PM

Hi!

I am trying to creat a research plan or more like designing my experiment.

I want to cut the DNA fragment with my gene of interest. I want to ligate it with a vector. I need the DNA fragment with restriction sites that I can use to digest the vector and then ligate them together. How can i do that? I mean, how can i add restriction sites to the DNA frgament with my gene of interest,

Thanks.

#2 Papaver

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Posted 11 October 2011 - 01:12 AM

You have to PCR-amplify your gene of interest using primers which contain the restriction site as an overhanging part of the primer. Then you can digest your PCR product and ligate it into the vector.

#3 minerva_29

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Posted 12 October 2011 - 05:10 AM

Papaver,

Thanks a lot!





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