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generate multiple cloning site in large vector


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#1 eve8

eve8

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Posted 27 August 2009 - 08:40 AM

Hi there,

I would like to insert a multiple cloning site into a large (>10kb) lentiviral vector before starting to clone everything blunt and maybe getting mad while trying ;) . I know that there are different options to do that, for example by adding restriction sites by PCR, or by annealing oligos with the desired sites and ligating them into the vector. Can anyone describe a detailed method which worked well for him an may be applicable for my large vector?

Thanks in advance




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