I am trying to clone two fragments into my vector of interest pcdna5 frt to.

I have insertedone fragment into the vector using cut and paste method.

Now i want to insert another fragment into the vector.i am trying to use gibson cloning in this step. Because the traditional method will not be suitable as maximum of restriction sites in the vector are also present in my fragment of interest.

But the results are not.positive ...i have tried chnging the vector insert ratio but only colonies without the insert are obtained. please suggest a way out or a methos to achieve my goal.

Thank you