Biology Forum Molecular Biology Cloning vector

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    • #9399
      roniadam
      Participant

      Hi everyone:

      how many enzymes are required to open a cloning vector for insertion of the cloned gene? I am using PFastBac vector and I cut it by one enzyme only. Is this will allow the interested fragment to insert into the vector or I have to use two enzymes???

    • #83219
      canalon
      Participant

      1 enzyme is enough but the risk is religation of the vector without integration of the fragment. And of course you wil not be able to select the orientation of your insert, which can be a problem if you plan to express a protein.

    • #87551
      camzie
      Participant

      i have a question:

      Why isn’t the plasmid that contains the strongest promoter always the best expression vectors?

    • #87708
      MrMistery
      Participant

      you might want to turn the expression on and off using an inducer…

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