Preparation of M63 Minimal Medium

OVERVIEW

M63 Minimal Medium is a chemically-defined growth medium. It  can be used to culture Escherichia coli. It is prepared as a concentrated medium without supplement which is diluted to a working solution and supplemented with appropriate carbon and energy source, and some trace elements.

The following section describes a procedure to prepare 1000 ml of M63 minimal medium.

REQUIREMENTS

Reagents and solutions
5x M63 Minimal Medium (without supplements)
20% Glycerol
1 M Magnesium sulfate (MgSO4)
Deionized water

Optional
0.5% vitamin B1 (thiamine)
20% Casamino Acids
Antibiotic

Equipment and disposables
Measuring cylinder
Conical flask / Beaker
Magnetic stirrer

Composition of M63 Minimal medium
1x M63 Minimal salts
0.2% Glycerol
1 mM MgSO4

Objective
Preparation of 1000 ml of M63 Minimal medium

PROCEDURE

Step 1: To prepare 1000 ml of M63 Minimal medium, take 700 ml water in a 2-liter beaker. Add 200 ml 5x concentrated M63 medium (without supplements), 10 ml of 20% Glycerol, and 1 ml of 1M MgSO4. Mix.

Note: if required, 0.1 ml 0.5% vitamin B1 (thiamine), 5 ml 20% Casamino Acids, and antibiotics can be added.

Step 2: Adjust the volume to 1000 ml with deionized water. Mix it again.

Step 3: Sterilize the solution by filtering through a filter unit containing a membrane with pore size 0.22 or less under aseptic conditions.

Tip:
You don’t need to filter-sterilize the solution if you have used sterile reagents and glasswares and handled them aseptically.

Storage:
Store the solution in fridge (2 – 8°C)

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