IGEM:PennState/2006/Growthmedia: Difference between revisions
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*Cool in such a way as to avoid accumulation of condensation, while maintaining sterility | *Cool in such a way as to avoid accumulation of condensation, while maintaining sterility | ||
*Seal with parafilm, place in bag, and store at 4ºC | *Seal with parafilm, place in bag, and store at 4ºC | ||
<h4>Top Agar (per liter)</h4> | |||
*500 mL LBM | |||
*500 mL DI H<sub>2</sub>O | |||
*8 g Bacto-Agar | |||
*Autoclave | |||
<h2>SOB</h2> | <h2>SOB</h2> | ||
| Line 52: | Line 57: | ||
*Add filter-sterilized glucose to SOB for 20 mM final concentration of glucose | *Add filter-sterilized glucose to SOB for 20 mM final concentration of glucose | ||
<h2>Minimal Media</h2> | |||
<h3>M9 Salts (per liter)</h3> | |||
* 1. Make M9 salts | |||
<h2> | * 2. To make M9 Salts aliquot 800ml H2O and add | ||
( | **64g Na2HPO4-7H2O | ||
**15g KH2PO4 | |||
* | **2.5g NaCl | ||
* | **5.0g NH4Cl | ||
* | **Stir until dissolved | ||
* | **Adjust to 1000ml with distilled H2O | ||
* | **Sterilize by autoclaving | ||
* | <h4>Media (per liter)</h4> | ||
< | * 3. Measure ~700ml of distilled H2O (sterile) | ||
* 4. Add 200ml of M9 salts | |||
* 5. Add 2ml of 1M MgSO4 (sterile) | |||
* 6. Add 20 ml of 20% glucose (or other carbon source) | |||
* | * 7. Add 100ul of 1M CaCl2 (sterile) | ||
* | * 8. Adjust to 1000ml with distilled H2O | ||
* | |||
* | |||
* | |||
Latest revision as of 06:39, 1 July 2008
Growth Media
LB
Media (per liter)
- 10 g Bactotryptone
- 5 g Yeast Extract
- 10 g NaCl
- pH to 7.0 with NaOH/HCl
- Autoclave
Plates
- Add 15 g Bacto-Agar per liter media
- Autoclave for 25 minutes
- If antibiotics necessary, add only once media has cooled to ~45ºC, or is comfortable to the touch
- Pour in sterile environment (~15ml per plate)
- Cool in such a way as to avoid accumulation of condensation, while maintaining sterility
- Seal with parafilm, place in bag, and store at 4ºC
LBM
Media (per liter)
- 10 g Bactotryptone
- 5 g Yeast Extract
- 5 g NaCl
- 0.4066 g MgCl2.6H2O (or 2mL 1M MgCl2)
- pH to 7.0 with NaOH/HCl
- Autoclave
Plates
- Add 15 g Bacto-Agar per liter media
- Autoclave for 25 minutes
- If antibiotics necessary, add only once media has cooled to ~45ºC, or is comfortable to the touch
- Pour in sterile environment (~15ml per plate)
- Cool in such a way as to avoid accumulation of condensation, while maintaining sterility
- Seal with parafilm, place in bag, and store at 4ºC
Top Agar (per liter)
- 500 mL LBM
- 500 mL DI H2O
- 8 g Bacto-Agar
- Autoclave
SOB
Media (per liter)
- 20 g bactotryptone
- 5 g bacto-yeast extract
- 0.5 g NaCl
- 0.19 g KCl
- pH to 7.0 with NaOH/HCl
- Autoclave for 25 minutes
- Add filter-sterilized MgCl2 and MgSO4 to yield final concentration of 10 mM each
Final Concentration:
- 0.5% Yeast Extract
- 2% Tryptone
- 10mM NaCl
- 2.5 mM KCl
- 10 mM MgCl2
- 10 mM MgSO4
SOC
Media
- Add filter-sterilized glucose to SOB for 20 mM final concentration of glucose
Minimal Media
M9 Salts (per liter)
- 1. Make M9 salts
- 2. To make M9 Salts aliquot 800ml H2O and add
- 64g Na2HPO4-7H2O
- 15g KH2PO4
- 2.5g NaCl
- 5.0g NH4Cl
- Stir until dissolved
- Adjust to 1000ml with distilled H2O
- Sterilize by autoclaving
Media (per liter)
- 3. Measure ~700ml of distilled H2O (sterile)
- 4. Add 200ml of M9 salts
- 5. Add 2ml of 1M MgSO4 (sterile)
- 6. Add 20 ml of 20% glucose (or other carbon source)
- 7. Add 100ul of 1M CaCl2 (sterile)
- 8. Adjust to 1000ml with distilled H2O
