STANDARD RECIPE
1 of 2
Latest update: 12/02/2018
RECIPE FOR THE PREPARATION OF COMPONENTS OF AUTOINDUCTION MEDIA
This protocol is based on the paper by William F Studier:
Studier (2005). Protein production by auto-induction in high-density shaking cultures. Protein
Expression and Purification vol. 41(1), p. 207-234
The protocol is slightly adapted from the paper. ZYM-5052 is the medium recommended for
standard auto-induction experiments.
ZY MEDIUM
MM (g mol-1)
Component
Tryptone (%)
Yeast extract (%)
Preparation
To be weighed in 1
liter (g)
To be weighed in 10
liter (g)
1 n.a.
10
100
0,5 n.a.
5
50
dissolve all components in the appropriate volume of MilliQ water (0.25 ml medium per
bottle)
Pour 0.25 ml medium in a 1 liter baffled culturing flask
Cover each flask with bottle cap and aluminium foil
Autoclave
50x M
Component
end
concentration
(mM)
MM (g mol-1)
stock
concentration
(M)
To be weighed
in 0.8 liter (g)
To be weighed
in 1 liter (g)
Na2HPO4
25
1,25
177,99
177,99
222,49
KH2PO4
25
1,25
136,08
136,08
170,10
NH4Cl
50
2,5
53,49
106,98
133,73
5
0,25
142,04
28,41
35,51
Na2SO4
Preparation
dissolve all components in the appropriate volume of MilliQ water (800 ml is the preferred
option)
Autoclave (do not firmly close the bottle!)
STANDARD RECIPE
2 of 2
Latest update: 12/02/2018
50x Mg
Component
end
concentration
(mM)
MgSO4
Preparation
MM (g mol-1)
stock
concentration
(M)
2
0,1
To be weighed
in 0.8 liter (g)
246,5
To be weighed
in 1 liter (g)
19,72
24,65
dissolve all components in the appropriate volume of MilliQ water (800 ml is the preferred
option)
Autoclave (do not firmly close the bottle!)
50x Fe
Component
Iron salt
end
concentration
(mM)
MM (g mol-1)
stock
concentration
(M)
0,01
0,0005
To be weighed
in 0.8 liter (g)
270,30
To be weighed
in 1 liter (g)
0,1081
0,1352
Fe(III)Cl3 6H2O is the iron salt used in this lab
Preparation
dissolve all components in the appropriate volume of MilliQ water (800 ml is the preferred
option)
Autoclave (do not firmly close the bottle!)
50x 5052
Component
end
concentration
(%)
stock
concentration
(%)
MM (g mol-1)
To be weighed
in 1 liter (g)
To be added in 1
liter (ml)
Glycerol (%)
0,5
25 n.a.
0
Glucose (%)
0,05
2,5 n.a.
25
0,2
10 n.a.
100
α-Lactose (%)
Preparation
250
dissolve all components in MilliQ water (heat to promote dissolving)
Filter-sterilise into 50 ml conical tubes using a 0.22 µm filter. Firmly close the tubes and cover
with parafilm