I) Had to re-run gel, it was wierd.
II) transformation of NEB10b and DH5a with the pcr product and ligation products.
III) Ligation with blunt end, cohesive end from Bangalore Genei and another cohesive end from Promega.
This will probably eliminate ligase related problems.
IV) Kan reduced from 50ul/ml to 25ul/ml
V) PCR to re-amplify test DNA,
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