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I have two inserts that I want to insert into a kb vector. What should be the insert: vector ratio?

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5.6-6.8 kb for vector

1.4-3 kb for inserts

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  1. 10 to 1 molar ratio (insert to vector).  Assuming your vector has a selectable marker and you dephosphorylate the vector with SAP (or something like that).    What restriction site are you using and why not do this in two steps, first one insert, then the next?

    It's been a while since I did this.  (more that 15 yrs) so my answer might be a little dated.

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