E. coli for 3.2

Submitted by kdorfman on Tue, 10/18/2011 - 15:18

GFP E. coli grown in LB

Grow cells in LB-Kan 1

Overnight culture

Dilute and monitor in morning; keep cells in active growing phase. Turn off heat in shaker if necessary.

Need ~20 mL total (8x12x2x100uL)

OD of culture should be 0.4 when cells are added to plates. That will put the OD just down into the noise, so there will be a lag time at the beginning of the run.

TA aliquots 100 uL cells to student wells, so the OD in the well will be half of the OD in the culture.

100 µL in 96 wells on 2 plates = ~20 mL of cells

Run at RT to be comparable to growth curves (set thermostat to 25C).

Use Script Mode on the plate reader. Program is called E_coli_glow&grow.

Check layout - make sure all rows used are read!

Fix file names so OD, FL, ps1, ps2 are obvious.

Move all the files into a folder labeled with the lab day as soon as the program is through running - otherwise, the data files for the second lab day are interleaved with those for the first lab day. (Sorting by time created may help you out of this fix, though.)

The 12 minute pause is to make the cycle take 15 minutes. Check that lab manual explains this. Enter the time on the compiled file in 15 minute intervals.

  1. Consider growing cells in LB+glu, then spinning down and resuspending in LB. They are slightly fluorescent in LB. ↩︎