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pJET1.2 ligation question

cloning pjet ligation

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#1 Druss00



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Posted 11 October 2013 - 09:59 AM



I have a quick question about pJET1.2 that I could use some help in.  I know it is linearized and phosphorylated at the 5' so why doesn't it re-ligate back together immediately when ligase is added?  I know it distrupts a lethal gene, so I would think that if someone tried to insert a blunt PCR product, pJET would ligate to itself (energy favorable) first and not allow insert a chance to "get in there".  Thus creating a lethal product.  I know this doesn't happen because we use it in lab, but I was curious.


Any insights would be appreciated!



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