IGEM:MIT/2007/Notebook/2007-7-14

Late Friday (7/13) Afternoon Update

• BH/SK put dry pT040 (?) plates in warm room (for AK and AL)
• Tom stopped by -- we have a BUNCH of chemically competent cells in the -80 degree freezer AND vectors pSB3AK3, pSB3AC3 and pSB3AT3 (already cut with E and P!) in the -20 degree iGEM freezer for our use -- y'all missed some fun with dry ice =)

Weekend Update

So Bernice and I (SK) came in this afternoon. Here's a quick summary of what happened:

• Results of testing for antibiotic resistance of parts

Part          Kan+         Amp+
F2620      no growth      growth
B0034      no growth      growth
E1010      growth        no growth

This is what we wanted to happen, so yay!

• Results of transformations (AK/AL)
  • Sample 1, 100 microliters and Sample 2, 100 microliters did not grow at all on A+/K+ plates

Sample          Volume              Number of Colonies
1               50 microliters      0
1               100 mics            4 or 5
1               200 mics            about 10
2               50 mics             2
2               100 mics            5
2               200 mics            10 to 12   

These plates should be parafilmed and in the 4 degree fridge.

• Made permanent glycerol stock of transformed XL1 cells after 3A transformation (in -80 degree freezer)
• Made LC of AK/AL plates using Sample 1, 200 microliters (2 colonies) and Sample 2, 200 microliters (2 colonies) -- they're in the rotating shaker in the hot room (put in around half 4 on Saturday)
• Miniprepped the diluted transformations of XL-1 cells after 3A transformation (so these have F2620 and B0034 in pSB3K3) -- from LC 1 and 2
  • The miniprepping we actually did twice because the nanodrop numbers were really, really bad the first time. From the second miniprep:

LC 1A = 36.9 ng/microliter
LC 1B = 32.7
LC 2A = 27.0
LC 2B = 24.2

These are all in the -20 degree iGEM freezer in Eppendorf tubes.