ATCC Medium: 158 RGCA Medium
Complete Medium
Solution 1 (see below)……………………….150 ml
Solution 2 (see below)……………………….150 ml
Solution 3 (see below)……………………….1 ml
Solution 4 (see below)……………………….220 ml
Rumen Fluid…………….……………………400 ml
Agar (if needed)………………………………15 g
Autoclave at 121?C. Allow to cool while stirring under CO2 gas. Aseptically add:
Solution 5 (see below)……………………….17 ml
Solution 6 (see below)……………………….66 ml
After continued gassing for 15 minutes, adjust pH to 6.6. Dispense aseptically into sterile gassed out tubes.
Solution 1
K2HPO4………………………………………….1.5 g
DI Water…………………………………………500 ml
Solution 2
KH2PO4………………………………………….1.5 g
(NH4)2SO4……………………………………….3.0 g
NaCl……………………………………………..3.0 g
MgSO4…………………………………………...0.3 g
CaCl2…………………………………………….0.3 g
DI Water…………………………………………500 ml
Solution 3
Resazurin……………………………………….0.05 g
DI Water…………………………………………50 ml
Solution 4
Glucose…………………………………………3.0 g
Cellobiose………………………………………3.0 g
DI Water…………………………………………325 ml
Solution 5
CysteineHCl…………………………………….0.75 g
DI Water………………………………………….25 ml
Boil briefly under CO2 gas. Seal and autoclave at 121?C.
Solution 6
Na2CO3………………………………………………6.0 g
DI Water……………………………………………...100 ml
Boil briefly under CO2 gas. Seal and autoclave at 121?C.