Magni:Protocols/M9: Difference between revisions
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== CaCl2 1M == | == CaCl2 1M == | ||
*Dissolve powder in approximately 80 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer. | *Dissolve powder in approximately 80 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer. | ||
*Bring to a volume of 100 ml. | *Bring to a volume of 100 ml. | ||
| Line 48: | Line 42: | ||
*Autoclave in a pyrex bottle. | *Autoclave in a pyrex bottle. | ||
*Store at room temperature and use near the flame. | *Store at room temperature and use near the flame. | ||
== MgSO4 1M == | |||
*Dissolve powder in approximately 80 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer. | |||
*Bring to a volume of 100 ml. | |||
*Autoclave in a pyrex bottle. | |||
*Store in the fridge and use at room temperature, near the flame. | |||
</div> | </div> | ||
Latest revision as of 05:43, 11 December 2012
M9 supplemented medium (liquid)
The final composition of the medium is: M9 salts, 1 mM thiamine hydrochloride, 0.2% casamino acids, 2 mM MgSO4, 0.1 mM CaCl2, 0.4% glycerol.
- For every 1 L of medium, add to a bottle or flask (in the following order) while using a sterile magnetic stirrer:
- 739 ml of autoclaved deionized water (blue tap of the Millipore machine)
- 100 ul of CaCl2 1M
- 200 ml of 5X M9 salts
- 20 ml of casamino acids 10%
- 34 ml of thiamine hydrochloride 10 mg/ml
- 5 ml of glycerol 80%
- 2 ml of MgSO4 1M
- Store in the fridge. NOTE: medium occasionally precipitates when stored for long periods.
CaCl2 1M
- Dissolve powder in approximately 80 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer.
- Bring to a volume of 100 ml.
- Autoclave in a pyrex bottle.
- Store in the fridge and use at room temperature, near the flame.
5X M9 salts
- Dissolve 56.4 g of powder in approximately 800 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer.
- Bring to a volume of 1 L.
- Autoclave in a pyrex bottle.
- Store in the fridge and use at room temperature, near the flame.
Casamino acids 10%
- Dissolve 10 g of powder in approximately 80 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer (the solution will not become clear before autoclaving).
- Bring to a volume of 100 ml.
- Autoclave in a pyrex bottle.
- Store in the fridge and use near the flame.
Thiamine hydrochloride 10 mg/ml
- Dissolve 0.4 g of powder in exactly 40 ml of Milli-Q water (from the filter of the Millipore machine) in a 50-ml tube.
- Filter-sterilize with a 20-ml syringe and a 0.2 um filter.
- Do not store! use it when needed to prepare the medium.
Glycerol 80%
- Mix 80 ml of glycerol with 20 ml of deionized water (blue tap of the Millipore machine).
- Autoclave in a pyrex bottle.
- Store at room temperature and use near the flame.
MgSO4 1M
- Dissolve powder in approximately 80 ml of deionized water (blue tap of the Millipore machine) by using a magnetic stirrer.
- Bring to a volume of 100 ml.
- Autoclave in a pyrex bottle.
- Store in the fridge and use at room temperature, near the flame.
