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How many copies of the construct get into bacteria during transformation?

How many copies of the construct get into bacteria during transformation? - Molecular Cloning Forum

How many copies of the construct get into bacteria during transformation? - 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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Old 05-22-2010, 08:10 PM
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Default How many copies of the construct get into bacteria during transformation?



Hello, guys,

I would appreciate help here. I would like to ask if anybody knows how many copies of your construct are "engulfed" by bacteria during transformation? I use DH5alpha, I dont know whether it matters. I could not find info on that. Thank You!!
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Old 06-02-2010, 05:24 PM
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Default Re: How many copies of the construct get into bacteria during transformation?

Well, the number of plasmid a bacteria can get during a transformation depends on various factors, mainly the efficiency of the competent cells. DH5alpha is an E. coli strain used for transformations with good results, but DH5alpha itself is not transformable without a process to make it competent.

It will also depend on whether the plasmid you have is supercoiled or not, since it's more easily engulfed a supercoiled plasmid than a relaxed one.

It will depend also on the size of the construction. Smaller vectors are more easily engulfed than larger ones.

It will depend also on which process you use to transform. Heat-shock transformation yields different results than Electroporation.

Anyway, I don't know why do you wanna calculate how many copies of your vector will a bacteria engulf, because when inside the bacteria, it will replicate and the number will encrease.

Hope it helps
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Old 06-18-2012, 06:53 AM
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Default Re: How many copies of the construct get into bacteria during transformation?

thanks
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Old 06-21-2012, 05:22 PM
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Default Re: How many copies of the construct get into bacteria during transformation?

It seems to me based on transformation efficiencies that you get 1 copy in. But as said earlier, this is then copied many times.
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