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on the minimal pcr derived product


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#1 Roberto

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Posted 13 May 2009 - 09:06 AM

Hello,
What would be the minimal length for a PCR derived product to be handled (plasmid cloned?)
Many thanks.

#2 mastermi

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Posted 13 May 2009 - 12:05 PM

What do you mean?
You want to amplify a very short product and clone it into a plasmid?

You can clone nearly everything, of course it gets harder to handle the more short the product is, because you lose more DNA during purification steps etc.
The minimal length of a DNA fragment I was working with was 35 bp. But I didn't PCR-amplify it but used complement oligos to create my dsDNA.

#3 Roberto

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Posted 13 May 2009 - 01:22 PM

What do you mean?
You want to amplify a very short product and clone it into a plasmid?

You can clone nearly everything, of course it gets harder to handle the more short the product is, because you lose more DNA during purification steps etc.
The minimal length of a DNA fragment I was working with was 35 bp. But I didn't PCR-amplify it but used complement oligos to create my dsDNA.



Many thanks for your comment. I have a plasmid clone, from which I want to subclone a 70b region, I was wondering if such a small fragment could be handled to PCR it and subclone it. I was considering using a Topo vector. From your comment I can go ahead... Many thanks again.
Roberto

Edited by Roberto, 13 May 2009 - 01:23 PM.





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