Hi,
I am trying to calculate the efficiency of transformation of my electrocompetent cells. To do that, I calculated the average of the colonies I obtained on tree plates streaked. I was wondering if it normal that the number of colonies vary a lot among the different plates, or it is supposed to be more or less the same..In 1 plate I counted 1160 colonies, in another one 560 and in the third one 250..Does this happen? To what is it due?
Thanks
The variation could be due to: (1) The amount of competent cells you removed/used could be different, (2) the plasmid amount you removed/used for transformation could be different too, plus (3) the amount of electroporated cells you streaked on the plates could be different. (4) the unfavorable conditions (such as air bubbles, which might cause sparks) might cause the decrease of transformation efficiency.
Since you never can reproduce every single condition (amount of cells, transfacted cells etc.) you always will get variation in your colony amount.
The height of your agar plate (and therefore the concentration of your antibiotic) is also a factor to consider.
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