Protocol for Culturing Streptavidin:
Preparation of Materials:
Basic Salts and Trace Elements (TE+BS):
Salt | Amount |
---|---|
H3BO3 | 500 ug |
CuSO4 5H2O | 40 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