Transformation frequency calculation

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medmagic
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Transformation frequency calculation

Post by 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

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canalon
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Post by 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

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

amoyensis
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Re: Transformation frequency calculation

Post by 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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