Magni:Protocols/M9 - Revision history

Lorenzo Pasotti at 12:43, 11 December 2012

2012-12-11T12:43:20Z

←Older revision		Revision as of 12:43, 11 December 2012
Line 16:		Line 16:

	== CaCl2 1M ==		== 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.
-
-	~~== MgSO4 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>

Lorenzo Pasotti at 12:42, 11 December 2012

2012-12-11T12:42:43Z

←Older revision		Revision as of 12:42, 11 December 2012
Line 16:		Line 16:

	== CaCl2 1M ==		== 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.
		+
		+	== MgSO4 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.

Lorenzo Pasotti: /* M9 supplemented medium (liquid) */

2012-07-31T18:22:55Z

M9 supplemented medium (liquid)

←Older revision		Revision as of 18:22, 31 July 2012
Line 13:		Line 13:
	**5 ml of glycerol 80%		**5 ml of glycerol 80%
	**2 ml of MgSO4 1M		**2 ml of MgSO4 1M
		+	*Store in the fridge. NOTE: medium occasionally precipitates when stored for long periods.

	== CaCl2 1M ==		== CaCl2 1M ==

Lorenzo Pasotti: /* M9 supplemented medium (liquid) */

2012-07-31T18:21:17Z

M9 supplemented medium (liquid)

←Older revision		Revision as of 18:21, 31 July 2012
Line 4:		Line 4:
	= M9 supplemented medium (liquid) =		= 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.		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:		*For every 1 L of medium, add to a bottle or flask (in the following order) while using a sterile magnetic stirrer:

Lorenzo Pasotti: New page: {{Magni}}
= M9 supplemented medium (liquid) = The final composition of the medium is: M9 salts, 1 mM thiamine hydroch...

2012-07-31T18:21:00Z

New page: {{Magni}} <div style="padding: 10px; width: 720px; border: 5px solid #000000;"> = M9 supplemented medium (liquid) = The final composition of the medium is: M9 salts, 1 mM thiamine hydroch...

New page

{{Magni}}

<div style="padding: 10px; width: 720px; border: 5px solid #000000;">
= 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

== 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.

</div>

Magni:Protocols/M9 - Revision history

Lorenzo Pasotti at 12:43, 11 December 2012

Lorenzo Pasotti at 12:42, 11 December 2012

Lorenzo Pasotti: /* M9 supplemented medium (liquid) */

Lorenzo Pasotti: /* M9 supplemented medium (liquid) */

Lorenzo Pasotti: New page: {{Magni}} = M9 supplemented medium (liquid) = The final composition of the medium is: M9 salts, 1 mM thiamine hydroch...

Lorenzo Pasotti: New page: {{Magni}}
= M9 supplemented medium (liquid) = The final composition of the medium is: M9 salts, 1 mM thiamine hydroch...