help with transformation efficiency calculation! - (Apr/20/2010 )
Hi
im having trouble calculating the transformation efficiency for the competent cells. this is what i was given:
5 �l pUC18 (0.1 ng/ �l)
50 �l E. coli competent cells
Left on ice 30 minutes
Heat shocked at 42C for 45 seconds
Added 500 �l NZY broth
Place in 15 ml culture tube in shaking incubator 37C for 1 hr
Then spread 10 �l, 50 �l and 100 �l on different LB amp plates.
10 �l gave 495 colonies.
would really appreciate it if someone could help me out, the answer should be in cfg/ug units
thanks!
-sc7-
-sc7-
sc7 on Apr 20 2010, 04:51 PM said:
Hi
im having trouble calculating the transformation efficiency for the competent cells. this is what i was given:
5 �l pUC18 (0.1 ng/ �l)
50 �l E. coli competent cells
Left on ice 30 minutes
Heat shocked at 42C for 45 seconds
Added 500 �l NZY broth
Place in 15 ml culture tube in shaking incubator 37C for 1 hr
Then spread 10 �l, 50 �l and 100 �l on different LB amp plates.
10 �l gave 495 colonies.
would really appreciate it if someone could help me out, the answer should be in cfg/ug units
thanks!
-sc7-
im having trouble calculating the transformation efficiency for the competent cells. this is what i was given:
5 �l pUC18 (0.1 ng/ �l)
50 �l E. coli competent cells
Left on ice 30 minutes
Heat shocked at 42C for 45 seconds
Added 500 �l NZY broth
Place in 15 ml culture tube in shaking incubator 37C for 1 hr
Then spread 10 �l, 50 �l and 100 �l on different LB amp plates.
10 �l gave 495 colonies.
would really appreciate it if someone could help me out, the answer should be in cfg/ug units
thanks!
-sc7-
This sounds like a homework question. You should probably go to the homework question forum if that is the case.
Anyway, the key is to calculate how much DNA (in �g unit) you had in that 10�l you plated, which is not difficult.
And I believe the unit should be cfu/�g, in which cfu means Colony Forming Unit.
-ProteinWork-