Dandekar & Chandler:Selective media
Nicole Smalley (Talk | contribs)
(New page: Back Home: Dandekar & Chandler:Protocols ==Selective Media== ===Selective Antibiotics=== *Trimethoprim (for pseudomonas selection) ::Working concentration [100 ug/ml] from DMSO stock [...)
Next diff →
Revision as of 18:08, 22 August 2013
Back Home: Dandekar & Chandler:Protocols
- Trimethoprim (for pseudomonas selection)
- Working concentration [100 ug/ml] from DMSO stock [100 mg/ml]
- Triple antibiotic (for burkholderia selection from co-culture)
- 150,000 IUs/L Polymixin B (=17.3 mg/L)
- 5 mg/L gentamicin
- 100 mg/L ticarcillin
Useful for selection of LasR mutants from WT Pseudomonas aeruginosa
After competition against another species, first spread dilute co-culture onto antibiotic plate, then patch isolated pseudomonas onto milk plates.
WT colonies will create clearings, allowing determination from LasR mutants.
Skim Milk Plates
Do not combine LB and milk solutions before autoclaving -- they react and cause a giant mess!
1. Prepare the following solutions:
- 150 ml H2O
- 150 ml LB (can be previously autoclaved)
- 9g bacto-agar
Stir and autoclave; keep molten!
- 300 ml H2O
- 24 g powdered milk (2%)
Stir well, leave stir bar in flask and autoclave
2. Autoclave solutions separately
3. Start milk solution stirring on stir plate
4. Slowly pour molten LB agar into milk solution
Recipe above pours one sleeve of plates