Why did some Colonies Grow?

darkwng · #1 (**permalink**) 05-19-2008, 04:37 PM

Why did some Colonies Grow?

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

any ideas?

kmunson779 · #2 (**permalink**) 05-19-2008, 06:19 PM

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?)

Andriy · #3 (**permalink**) 07-01-2008, 09:46 PM

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

trypsin · #4 (**permalink**) 07-07-2008, 06:42 PM

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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Why did some Colonies Grow?

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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.