20.109(S11): TA notes for module 2
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Revision as of 14:53, 8 March 2011 by AgiStachowiak (Talk)
General notes
Key preparation:
Scheme:
Day-by-day
Day 1
NOT YET COMPLETE - SEE ALSO GOOGLE DOC
Materials required:
- Two days before lab, streak out the following strains:
- ABX(8?) = pEDL3/pCph8/pPLPCB, on Amp/Cam/Kan plate
- ABX(22?) = pED-IPTG-INS, on Amp plate
- One day before lab, prepare O/N cultures of same
- AB8? is for edge detection plates
- AB22? is for first liquid culture experiment
Below are at set up at teaching bench unless otherwise noted:
- Equipment
- Both water baths
- Consumables
- Bags of 14 mL rb tubes (56 tubes needed per day)
- Pack of 50 mL conical tubes
- 2 boxes of cuvettes
- 5 and 10 mL pipets, pipet-aid
- Empty Petri dishes
- 15 mL conical tubes
- Photomasks
- Reagents (see google doc for amounts not listed)
- LB aliquots: ~30 mL per group (25 + for OD measurement + excess)
- On ice, antibiotic and additive aliquots, about 1 per 2 groups
- Plain ampicillin
- AHL
- IPTG
- Amp/Cam/Kan cocktail
Day of Lab (T/R):
- Prepare supplemented LB medium (30' autoclave, 30' or 60' for pressure to go down in large or small autoclave, respectively) and cool in a 42 °C water bath for at least 1 hour
- Turn on water bath so it has plenty of warm-up time, e.g. when begin autoclaving
After Lab
- Turn water bath back off.
How it went:
Day 2
Etc.
Special materials
Cell strains
- NB399 = JW3367c (EnvZ-, lacZ-, Kan^R removed)
- NB435/AB? = pEDL3, pCph8, pPL-PCB (full ED system)
- NB? = light to lacZ only
- AB? = pED-IPTG-110
- AB? = pED-IPTG-112
- AB? = pJT104 (full ED system less AHL)