Layne 9:30am
Yesterday's transformations:
P1010 Kan in ccdB strain - all plates showed some growth (a few small colonies)
P1010 Cm in ccdB strain - all plates showed some growth (a few small colonies)
Set 1
P1010 Kan 200ul - 5 (tiny)
P1010 Kan 100ul - 2 (tiny)
P1010 Cm 200ul - 7
P1010 Cm 100ul - 8
Set 2
P1010 Kan 200ul - 2
P1010 Kan 100ul - 1
P1010 Cm 200ul - 7
P1010 Cm 100ul - 2
I'm going to go ahead and assume this was a success. I'll pick from two each (Kan and Cm) of the best looking plates in about an hour or so and inoculate ne wbroth cultures. These results (small colonies overnight) aso indicate that our previous transformation may have been successful, although why we didn't get any plasmid on the gel we ran is still unknown.
Inoculated about 50ml of LB with two samples each of Kan and Cm at 11:30am.
Put into shaker at 37oC.
1:00pm - Kyliah
Well, I'll note that the ?failed P1010 plates didn't have colonies at 9 in the morning; it took until noon before they were even visible, and they were pinprick size all day. These colonies are actually decently sized.
3:10pm - Kyliah
Well, I took the OD450 of 1mL each of samples B (0.014) and C (0.063) - these are NOWHERE NEAR what they should be for log phase, so I'm not going to bother freezing any of them. Samples A and D look to be the same density to my eye. I will leave all four subcultures, and the two overnights, in for now.
3:40pm - Kyliah
I just sent off a parts request for I0500/pAra, R0051/pλcI, and all three of the P1010/ccdB parts (just in case, as we've not checked the amp or cm ones for plasmid yet).
4:05pm - Kyliah
Picked and inoculated C0051, B0015, B0034, J23066 for overnight culture (hopefully they still have plasmids... couldn't get into freezer for samples to plate). I know we didn't precisely need to re-culture if we already have plasmid samples, but what the heck, why not. It'll tell us if the plates are still viable samples. (Actually in the waterbath at about 5pm.)
I'll pre-setup tomorrow's lab notebook with plans right now
(back to Lab notebook)
Comments (1)
james jacoby said
at 10:06 am on Aug 18, 2009
Awesome! That's great news. I was just getting set to come in, but hearing this I may wait until this afternoon instead. If you'll stick them in the broth culture then I'll come in around 2 when they're at mid-log phase and cryopreserve them. Yay!
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