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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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Why did some Colonies Grow?

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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  #1 (permalink)  
Old 05-19-2008, 04:37 PM
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Angry Why did some Colonies Grow?

Why did some Colonies Grow?

i cant understand it! after ligation and plating i only get 2 colonies

any ideas?
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  #2 (permalink)  
Old 05-19-2008, 06:19 PM
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Default 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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Old 07-01-2008, 09:46 PM
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Default Re: Why did some Colonies Grow?

Besides, if any of those colonies contains the correct construct - what else do you want in life? :-)
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