Magni:Protocols/M9

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(M9 supplemented medium (liquid))
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.

Revision as of 07:42, 11 December 2012

Home        Contact        Lab Members        Publications        Research        Internal        Protocols        Data       


Contents

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.

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.

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.
Personal tools