Alipasha

Imani
Mr.karim
SBI3U
December 3

Way to Glow

Bacterial transformation is a process by which a bacterium expresses and takes up the foreign
genetic material. Before we start to work in the lab, we should obey the safety rules. Wear
Coat, Goggles, and Gloves all the time. Now we label two microcentrifuge tubes, “+DNA”
and “-DNA”. Then we use an F250 to add 250 µl of the CaCl2 solution to each tube.
After that, we use a sterile inoculating loop to pick colonies from the source plate of E. coli
cells and add it to two tubes that we have and suspend the cells completely by tapping.
To the tube labelled ‘+DNA,’ we add 10 µl of fluoro green (pFG), twice. Then we place both
transformation tubes at 42°C in the dry bath incubator for 90 secs. Then we use an F250
micropipette to add 250 µl of LB (Luria Bertani) Recovery Broth to each tube. Finally
Incubate the cells for 20 minutes at 37°C in the dry bath incubator for their recovery period.
Now we can plate the cells. We use an F250 micropipette to transfer recovered cells from the
tube and spread the cells with a sterile inoculating loop for both “-DNA” and “+DNA”.
After the cell suspension is completely absorbed by the agar, the educator will place
the plates in the inverted position in a 37°C incubation oven for overnight.
A transformation efficiency is a quantitative number that shows the extent to which you
genetically transformed E. coli cells in the experiment.

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