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Team:University of Chicago/Notebook/Damonwang
From 2008.igem.org
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Friday 2008-07-18
pUC8 CVX 336C/JM109 cultures: one is turbid and one is clear. Caulobacter plates are ready, will inoculate a starter culture. E. coli strain DW01 (pUC8 CVX 336C/JM109) frozen in 200uL 80% glycerol per 800uL culture (~16% glycerol final concentration), in Box A, slots 1 and 2. Caulobacter in 5mL and 50mL PYE, picked separately, incubated ~30C, 100rpm. Some medium dripped down the neck of the flask, but I flamed the area.
Saturday 2008-07-19
Couldn't get into the building to check temperature or OD600. Apparently Daniel Choi happened to run into someone leaving, and got in that way. The professor says to see Julie Caldwell about 24hr access.
Monday 2008-07-21
Julie Caldwell sent in a request to add me to the 24hr access list.
To do
- check OD600 on Caulobacter cultures; if ready, freeze glycerol stocks.
- Test E. coli glycerol stock DW01 (JM109/pUC8 CVX 336C) by streaking an LB-chloramphenicol plate
-
Pour PYE-Agar plates, 5 w/ and 5 w/o 2ug/mL chloramphenicol (CM).Have PYE plates, need to pour LB-CM plates. In fact, just make a liter each of LB and PYE. - Need to set up for making electrocompetent cells. Sterilize
- 250mL flask
- 2L flask
-
34x250mL centrifuge bottles - 2L water (MilliQ)
- 10% glycerol (in water? yes)
- 2x30mL Corex tubes
- 35x1.5mL Eppendorf
50mL Caulobacter culture is OD600=2.0, so overgrown. (Cecil, blanked with PYE)
New plan
- Seed 50mL Caulobacter culture for electrocompetent stocks
- Streak PYE plate from overgrown Caulobacter culture
- Nora will make PYE
- Pour LB-CM for streaking DW01 from glycerol
Tomorrow, Nora will dilute the 50mL culture and I will make electrocompetent stock. When the Caulobacter plate is ready, seed 50mL for glycerol stock
Nora's SOB media and my stuff for electrocompetent Caulobacter autoclaved (left autoclave, 15min fluid cycle (cycle 4?)---Schonbaum says with several liters of fluid, better to use the 30min fluid cycle).
All the glassware for electrocompetent Caulobacter autoclaved (right autoclave, cycle 2) and put on lower shelf under micropipetter tips.
Remind Jim and Soren that every label should have date and owner's name. Nora's media flasks just said "SOB".
Julie Caldwell sent in a request for my 24hr access.
Plated Caulobacter PYE and Caulobacter LB+CM. Oops.
Poured 6 LB+CM, now in 4C.
Should have autoclaved PYE in 250mL flask. Now just took a guess, since no sterile graduated cylinders available. Must wait for PYE to cool in cool water bath.
Caulobacter DW02 ~175rpm, 30C, PYE 50-75mL
E. coli DW01 streaked on LB-CM from glycerol.
Tuesday 22 July 2008
| Time | OD600 |
|---|---|
| 9:45 | .006 |
| 11:52 | .017 |
| 13;50 | .033 |
Will come in ~11pm to dilute Caulobacter to appropriate concentration for Nora tomorrow.
16:00 Starter culure from streaked DW01 plate from glycerol at 37C, 250rpm, 5mL LB+8.75uL 2mg/mL CM
17:00 Starter culture from Crosson's DW01 plate picked to 30C, ~170rpm, 5mL PYE
The plate streaked 21 July 2008 from vastly overgrown culture has solid paths of colony---not Caulobacter because that's too fast
22:50 Our keycards still don't work, but apparently someone came out just as Nora was trying to enter.
Wednesday 23 July 2008
Friday 25 July 2008
Monday 28 July 2008
Tuesday 29 July 2008
Thursday 31 July 2008
Appendixes
Naming System
- Cultures are named by starting date, species, medium, and a number. Only as many of these as will give a unique name, are used.
- Strains are numbered sequentially, prefixed by DW to indicate that it's mine, all mine!
- Maybe they should have a species field too. Except I'm not likely to have more than three or four strains ever.
Freezer Box Inventory
| Slot | Date frozen | Strain | Description |
|---|---|---|---|
| 1 | 18 July 2008 | DW01 | Crosson's E. coli JM109 carrying pUC8 CVX 336C |
