Re: Mutating CMV promoter

Posted by Thermus aquaticus on Jan 26, 2002 at 02:25

Re: Mutating CMV promoter (Miryan)

Hmmm, I see.

But why can’t you cut out your insert with NdeI and end-fill it with T4 DNA pol or Klenow? Nde leaves a 5’ overhang, and both T4 pol and Klenow have a 5’ – 3’ polymerase activity. By my reckoning this should end fill your insert and leave the ATG intact. Then you can blunt ligate your insert into any site in the MCS of your vector.

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