Difference between revisions of "20.109(S11): TA notes for module 2"
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'''Materials required:''' | '''Materials required:''' | ||
− | * Two days before lab, streak out the following strains: | + | * '''Two days before lab''', streak out the following strains: |
** ABX(8?) = pEDL3/pCph8/pPLPCB, on Amp/Cam/Kan plate | ** ABX(8?) = pEDL3/pCph8/pPLPCB, on Amp/Cam/Kan plate | ||
** ABX(22?) = pED-IPTG-INS, on Amp plate | ** ABX(22?) = pED-IPTG-INS, on Amp plate | ||
− | * One day before lab, prepare O/N cultures of same | + | * '''One day before lab''', prepare O/N cultures of same |
**AB8? is for edge detection plates | **AB8? is for edge detection plates | ||
**AB22? is for first liquid culture experiment | **AB22? is for first liquid culture experiment | ||
− | Below are at set up at teaching bench: | + | Below are at set up at teaching bench unless otherwise noted: |
− | * 2 boxes of cuvettes | + | *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):''' | '''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 | * 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 | ||
+ | * <font color=FF3300>Turn on water bath</font color> so it has plenty of warm-up time, e.g. when begin autoclaving | ||
'''After Lab''' | '''After Lab''' | ||
+ | |||
+ | *<font color=FF3300>Turn water bath back off.</font color> | ||
'''How it went:''' | '''How it went:''' |
Revision as of 14:53, 8 March 2011
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)