Team:Unviersity of Chicago/Notebook/Damonwang

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Contents

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

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
    • 3 4x250mL 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".

Appendixes

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