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| Molecular Cloning Forum DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here. |
Phosphorylating Oligos
Molecular Cloning Forum
DNA cloning forum. Discuss the cloning of plasmids, vectors, DNA, RNA, cDNA, proteins, and other molecules for expression, sub-cloning, and other methods. Also discuss digestion, ligation, transformation, plasmid preparation, cDNA library screening, and competent cells here.
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02-20-2008, 01:28 AM
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 Phosphorylating Oligos Heya dudes, i am in the process of phosphorylating some oligos using the enzyme PNK. I added the following:
However it seems my Phosphorylation protocol failed. All of cloning were were negative!  . What should i do?My ideas it may be the ATP gone old/bad? I was using ATP (100mM) from which was prepared as a solution from Roche. The new ATP comes as a powder, and I make 100mM solution by dissolving it in water? is this the problem? thnx in advance        |
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