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Consider that I cut my vector backbone with a blunt end cutter (EcoRV) and Iam not performing the alkaline phosphate treatment which will preserve the 5' phosphate on the vector. Is it necessary...
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I have cloned a gene for a 10 kDa bacterial protein in pET28a (to be expressed as a C terminal His tagged protein) and transformed into DH5a. I have confirmed the clone by restriction digestion...
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