Phytone peptone........................................................................4.0 g
Trypticase peptone.....................................................................1.0 g
MgSO4 x 7H2O…………………..................................................0.4 g
K2HPO4 x 3H2O.........................................................................1.2 g
KH2PO4…………………….........................................................1.0 g
Phenol red (0.2%).......................................................................10.0 ml
Hemin chloride (0.1%).................................................................10.0 ml
*Gelrite.........................................................................................8.0 g
8% L-glutamine, freebase (add aseptically; see note below)......100.0 ml
BSA Fraction V (add aseptically) (20% solution).........................15.0 ml
DI Water……................................................................................865.0 ml
Measure ingredients, except for bovine serum albumin (BSA) and glutamine, into the distilled water. Mix well and adjust pH to 6.8 +/- 0.2. Bring to a boil to dissolve components. Autoclave at 121?C. Prepare the glutamine and BSA solutions and filter-sterilize. Add to partially cooled base medium and dispense complete medium as required.
*Glutamic Acid (Sigma G1251) can be substituted for L-glutamine, freebase.
*Omit Gelrite when preparing a broth
GELRITE gellan gum (formerly known as PS-60 and S-60) is a new naturally derived, highly purified polysaccharide which displays several interesting properties, including self gelling. The suitability of GELRITE as an agar substitute was tested by evaluating the performance of several media selected from among those most commonly used in the isolation, identification, and enumeration of microorganisms in clinical laboratories.