hi, im currently trying to clone a Nucleoprotein (NP) gene ~1.5kb inside a pGBKT7 vector ~7.3kb. The NP gene contains 2 RE sites, both EcoRI and BamHI. Im using the clontech InFusion cloning strategy. After, PCR, the gene was purified and ligated with the linearized pGBKT7 vector.
However, after sequencing the vector, only 220bp of the 1.5kb NP gene was present.
CAN ANYONE PLZ HELP ME? i would really appreciate it!
I ve tried a couple of times, but the results are still the same
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only partial gene is cloned inside the vector
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