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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. |
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#2
05-19-2008, 06:19 PM
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 Re: Why did some Colonies Grow? First thing I would check is the competency of my competent cells (do you have a positive transformation control)? Without any details on your protocol there are so many steps that could have gone wrong I can't suggest anywhere else specifically to look. Can you give some more details (restriction enzymes, blunt vs. sticky-end, phosphatase treatment? DNA concentrations?)  |
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#3
07-01-2008, 09:46 PM
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| Re: Why did some Colonies Grow? Besides, if any of those colonies contains the correct construct - what else do you want in life? :-) |
| The Following User Says Thank You to Andriy For This Useful Post: | ||
pigfarmer (10-28-2009)
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#4
07-07-2008, 06:42 PM
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| Re: Why did some Colonies Grow? My colonies always grow even though they've never had the right construct. That is strange!!! sorry, i have no idea. I also have problems with the cloning voodoo thing good luck |
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#5
09-27-2009, 08:05 PM
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| Re: Why did some Colonies Grow? Trypsin - are you treating your vector with a phosphatase? |
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#6
10-26-2009, 07:25 PM
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| Re: Why did some Colonies Grow? trypsin.. if you could give a little more details , that would be helpful.. how do u digest your plasmid and insert ? one or two enzymes? do you check the digested fragments on agarose gel? to see if the restriction enzymes work well. |
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Why did some Colonies Grow? 
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