Transformation Efficiency

Transformation efficiency is a measure of the competency of a cell and its ability

to uptake exogenous DNA. Cells can be made competent by growing cells to log phase,
holding at 4
o
C, and washing away media with ! m" CaCl
#
$di%alent cation&. The
standard method of reporting transformation efficiency is'
Transformation (fficiency ) * transformants +g plasmid DNA
Therefore, to calculate this you simply need the number of transformants and the amount
of DNA that ga%e rise to those transformants. The number of transformants can be easily
determined simply by counting the number of colonies growing on selecti%e media $plus
amp plates&. The amount of DNA gi%ing rise to those colonies can be determined by
asking how much DNA was ,plated- on the plate you .ust counted. This can be
determined by knowing the starting concentration of your transforming DNA, the amount
you pipetted into your competent cells, the dilutions you accomplished along the way
toward plating that plate, and the %olume of culture you plated onto the plate. /et-s
follow the DNA concentration through the process of a typical transformation.
0ipet 1! / of a 1 ng+/ DNA solution into 1!! / of compent cells.
2ow much DNA did you apply to your competent cells3
1! / 4 1 ng+/ ) 1! ng DNA
5ncubate on ice one hour, heat shock, add media to a final %olume of 1 m/, incubate at
6!
o
C, 6! minutes to allow for expression of the amp resistance gene.
7hat is the concentration of our DNA at this point3
1! ng of DNA is now in a %olume of 1 m/. Therefore'
1! ng DNA+m/ is our current DNA concentration.
7e now ha%e transformed cells and can plate them out on plus amp plates. 8ince we
don-t know the concentration of these transformed cells, we will accomplish a serial
dilution and plate. /et-s perform 6 ten fold dilutions of our transformed cells and plate
!.1 m/ of each dilution onto a plate. Now follow the DNA concentrations through this
process. 7e know our DNA concentration in our competent cell mix was 1!ng+m/.
After one ten fold dilution that concentration would be 1 ng+m/. Another ten fold
dilution will make it !.1 ng+m/ and a subse9uent ten fold dilution would make it !.!1
ng+m/. 7e then plate from these tubes !.1 ml and incubate.
2ow much DNA was ,plated- onto each of these 6 plates3
0late 1 from our first 1! fold dilution'
The concentration of DNA in that tube was already determined to be
1ng+ml. 7e plated !.1 m/ onto that plate, therefore'
1ng+m/ 4 !.1m/ ) !.1 ng DNA applied to that plate.
0late #' !.1ng+m/ 4 !.1m/ ) !.!1 ng DNA
0late 6' !.!1ng+m/ 4 !.1m/ ) !.!!1 ng DNA
8uppose plate 1 yielded #!! colonies, plate # yielded #! colonies, and plate 6 yielded
# colonies. 7hich plate are you going to count3 0late #, right3
8o, what do we know now3 7e ha%e the number of transformants on the plate and the
amount of DNA that ga%e rise to those transformants.
#! transformants+!.!1 ng DNA.
The con%ention is to report this figure as transformants+g DNA so we must con%ert.
Transformation (fficiency ) #! transformants+!.!1ng DNA 4 1!!!ng+1g
) #. 4 1!
:
transformants+g plasmid DNA