Team:Newcastle/Labwork/15 October 2009

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Microscopy Test to characterize KinA

Today we tried to test our transformed colonies with IPTG and get the results with microscope. We had overnight cultures in MM medium as if we were going to transform them. Our overnight cultures were in 25 ml of MM competence medium. We prepared the tubes as below. For each of them we prepared a flask with IPTG and without IPTG. IPTG was added after an hour we prepared them.

1. ON Culture 1 + MM medium
2. ON Culture 2 + MM medium
3. ON culture from teh colony with halo
4. B. subtilis 168 WT + MM
5. B. subtilis transformed with gfp-rrnb + MM

Every 30 minutes we took samples and recorded the growth curve.

We did not have good results using microscopy because our ON cultures did not grow very well. There was not much bacteria in the samples. Tomorrow we will repeat the same experiment but this time we will grow them in LB.

We prepared the LB+CHL and LB+CHL+Em solutions and inoculated the cultures. For B. subtilis we use

0.3ug/ml for Em
5ug/ml for CHL

in LB solutions