cloning in duet vector

[Sibapriya Chaudhuri]

hi
i m tryin to clone 2 genes in pRSF duet vector. the first gene has been successfully cloned and even its expression level checked by induction. i was able to effectively screen the first clone
bur somehow i am unable to clone the second gene. I am restricting the and mcs wit 2 different enzymes-hence not using phosphatase. the vector is being digested properly as evident from the gel profile. but not gettin nay positive clones while tryin to screen the transformants after ligation
can anyone help?


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