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LR Clonase II Reaction Mix

xedeen
in Biochemistry and Molecular Biology

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I am currently fused my interest plasmid into pENTR3C. Indeed after affirmative the sequencing and frame reading from 1st Base company. It seems the fusion was successfully done.

The next step is fusion the Entry clone into pAd/CMV/V5/DEST. The size of destination vector is >10kb. Optional to linearized the destination vector. But, for this case, I am not linearized the destination vector. Following the protocol strictly, I can't get the transfomants with correct insert. What should I do??? Is there any other optional step should I do??

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