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Transformation frequency calculation

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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Transformation frequency calculation

Postby medmagic » Wed Nov 19, 2008 8:56 pm

Hi,
I was wondering how I would go about calculating transformation frequency in transformants/ug of plasmid DNA used. The data that I have are the colony counts on antibiotic plates, the volume and dilution factor of the transformed cell preparation that was plated, and the amount of plasmid DNA and untransformed cell preparation that was used in the transformation protocol.
Thanks for your help
medmagic
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Postby canalon » Thu Nov 20, 2008 9:50 pm

with the amount of DNA and the Molecular weight of the plasmid you can calculate how many plamid you had in your transformation mix.
You can calculate how much of your mix was plated (100µl out of 1ml of transformation mix for example) to be taken into account (i.e. in this case you would consider a factor 10 for the proportion of plated plasmids)
A transformation efficiency of 100% would be 1 colony per plasmid. You can do the calculations now.
Patrick

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Re: Transformation frequency calculation

Postby amoyensis » Fri Apr 24, 2009 3:54 pm

sorry...i still don't get ur idea...here's my problem..
i have ermm...my plasmid weight = 0.5 uL
colony count is 15
Molecular weight of my vector (pTrcHis2) = 1361.50 kDa
how shud i link them all together o0?
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