hey every body
here is what i am facing right now:
1- I have a pwo amplified fragment with a restriction site for XhoI at the 3' end(3790 bp)
2- I have a fragment cloned in pUC18 with XhoI restriction site at the 5' end and EcoRI restriction site at the 3' end(80 bp)
3- I have to ligate these two fragments using the XhoI site and clone the resulting fragment in a vector in a way that i don't lose the EcoRI restriction site so that eventually I could digest the fragment with a specific restriction enzyme( that has a site in the 5' end of the big fragment) along with XhoI and proceed to the next step.
how is it possible to avoid losing Ecori's restriction site?
tnx
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