Protocol for Culturing Streptavidin:
Preparation of Materials:
Basic Salts and Trace Elements (TE+BS):

Salt Amount
H3BO3500 ug
CuSO4 5H2O40 ug
KI 100 ug
FeCl3 6H2O 200 ug
MnSO4 H2O 400 ug
NaMoO4 200 ug
ZnSO4 7H2O 400 ug
KH2PO4 1.0g
MgSO4 7H2O 0.5 g
NaCl 0.1g
CaCl2 2H2O 0.1g

SLMA:

Element Amount
L-Asn 7.0 g
Dextrose 10.0g
TE+BS 1.7g
K2HPO4 1.0g
water 1L

SLMB:

Element Amount
L-Asn 7.0 g
Dextrose 10.0g
TE+BS 1.7g
KH2PO4 1.0g
K2HPO4 1.0g
MgSO4 7H2O 0.5g
NaCl 0.1g
CaCl2 2H2O 0.1g
water 1L

Procedure:
1) Introduce inoculums into flask (preferably 4L that has been washed and autoclaved) of SLMA on shaker for 200 rev/min @ RT

2) Incubate until heavily growth of balls
3) Spin cells down, discard supernatant and wash with PBS
4) Transfer to SLMB after 1 week in SLMA for optimal cell production

5) Spin cells down, take supernatant out and test on SDS page

6) Transfer to SLMA after 1 week in SLMB for SA production

7) Spin cells down, take supernatant out and test on SDS page

Estimated time for culture: 2.5 weeks

Protocol by Tricia Lin January 2009