NEW BRUNSWICK SCIENTIFIC BioFlo 415, Manual To Operations

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BioFlo 415
y
M1360-0050 User’s Guide
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The following section outlines step-by-step procedures for carrying out a benchtop
fermentation. Provided in a question and answer format, this discussion covers such topics
as which media formulation, tubing size, and concentration of various additives should be
used. It also addresses the preparation, autoclaving and clean-up procedures for the vessel
and accessories. While this example refers specifically to an E. coli fermentation in a
BioFlo 415, the information is generally applicable for any fermentation.
23.1 Media Formulation
Question: What kind of media should be used, and does it differ from media
used in shake flasks?
Answer:
The media used in shake flasks does differ from the standard media used in
a fermentation vessel. Shake flask media is generally of a much simpler composition.
LB Broth and Tryptic Soy Broth are standard shake flask media.
Here is an example of a more complex media used in a recombinant
E. coli
fermentation:
Chemical g/L
KH
2
PO
4
3.5
K
2
HPO
4
5.0
(NH
4
)
2
HPO
4
3.5
MgSO
4
.
7H
2
O 0.5
Glucose 5.0 (for fed batch)
30.0 (for batch)
Yeast Extract 5.0
Trace Metals 1.0 ml/L
Antifoam 0.5 ml/L
Trace metals formulation:
FeCl
3
1.6
CoCl
2
.
6H
2
O 0.2
CuCl
2
0.1
ZnCl
2
.
4H
2
O 0.2
NaMoO
4
0.2
H
3
BO
4
0.05
Hcl 10 ml
H
2
O to 1000 ml