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EDVO-Kit # 222 Transformation of E. coli with pFluoroGreen™ and pFluoroBlue™

15

Transformation of E.coli

13. After the recovery period. remove Ihe fubes from the water both and place Ihem on Ihe lab bench. Proceed to plating the cells for incubation.

PL ATI IMG THE CELLS

Plating cells from the tubę labeled DNA" (Contro! Experiment):

To avoid contaminacion when plating, do not set che lid down on the lab bench - lift the lid of the piąte only enough to allow spreading. Be careful to avoid gouging the loop into the agar.

14. Use a steriłe I ml pipet to transfer recovered cells from the tubę labeled ” -" to the middle of the foilcwing plates:

•    0.2 ml to the piąte labeled LB -

•    0.2 ml to the piąte

labcied LB / AMP -

15.    Spread the cells with a steriie inoculating loop. (See Figurę at right)

16.    Cover both plates and allow the liguid to be cbsorbed.

Experiment Procedures

W

Plating cells from the tubę labeled "+ DNA"

17.    Use a stcrile 1 ml pipet to transfer recovered cells from the tubc labeled ''+ DNA” to the middle of the following plates:

•    0.2 ml    lo the piąte labeled L3 + (Green. Blue or

both plcsmids)

• 0.2 ml    to the piąte labeled LB / Amp + (Green. Blue or

both plasmids)

18.    Spread the cells with a steriie inoculating loop in the same manner as step 15.

19.    Cover the piąte and allow the liguid to be absorbed.

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