I am currently having trouble during cloning 500bp~1000bp inserts into Dh5alpha, the inserts were pcr products and blunt end ligation with my vector, however after performing colony pcr, I hardly get anything above 500bp which I need to analyze, please can some one let me know is it becuase of the smear DNA is hard to clone or there is something I can do to increase the efficency, thx!!
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