Efficiency of the pGLO transformation.
Transformation efficiency=
plate
Total number of colonies (cells) growing on the agar
--------------------------------------------------------------------------Amount of DNA Spread on the agar plate
1. Count the total number of colonies = ________________
2. To determine the amount of pGLO DNA spread we must have two pieces of
information:
a. The total amount of DNA we began with. This is the product of the
concentration and the total volume used or
DNA (ug) = (conc. of DNA (ug/ul) X (volume of DNA in ul)
Total amount of DNA (ug) used = _________
b. The total fraction of pGLO DNA ( in the bacteria) that you actually spread
onto the LB/amp/ara plate. to do this use the following formula:
Fraction of DNA used = volume spread on plate
------------------------------total volume in the test tube
fraction of DNA = _______________
c.
pGlo DNA spread (ug) = total amt of DNA used X fraction of DNA
pGLO spread (ug = _______________
Transformation efficiency=
plate
Total number of colonies (cells) growing on the agar
-------------------------------------------------------------------------Amount of DNA Spread on the agar plate
___________ transformants/ug
This number describes how effective you were in getting a plasmid into the bacteria.
The general agreement for the protocol you just did has an efficiency of between 7.0 x
10 to the 3rd and 9.0 x 10 to the 3rd.