ATCC Medium: 1773 Hexamita Medium
TYGM Medium (ATCC Medium 1171, see below)………………………….250 ml
Sonneborn’s Paramecium Medium (ATCC Medium 0802, see below)………750 ml
For Test Tubes
Prepare ATCC Medium 1171 and ATCC Medium 0802 separately. After each medium
has been sterilized and has cooled, aseptically combine 3.0ml of ATCC Medium 1171
and 9.0 ml of ATCC Medium 802 in 16 x 125 mm screw-capped test tubes.
ATCC Medium 1171
K2HPO4 ................................................2.8 g
KH2PO4 ................................................0.4 g
Yeast Extract (BD 211928)………........3.0 g
NaCl........................................................7.5 g
Gastric mucin..........................................2.0 g
Distilled water........................................970.0 ml
Prepare ingredients in order listed. Autoclave for 15 minutes at 121?C.
Cool. Aseptically add:
Bovine serum (heat-inactivated)...............................................30.0 ml
10% Tween 80 in absolute ethanol (filter-sterilized)................0.5 ml
Sterile Rice Starch Suspension (0.15ml/tube, See Below)…...18.75 ml
Aseptically dispense 8.0 ml per 16 x 125 mm screw-capped test tube.
While dispensing, keep medium stirring so that rice starch is equally distributed to
each test tube.
Rice Starch Suspension
Heat-sterilize 0.5 g rice starch (MP Biomedicals, Catalog # 102955) at 150?C for
2 hours. After sterilization and just prior to use, aseptically add 9.5 ml sterile
phosphate buffered saline solution (pH 7.4). Extra rice starch solution should not
be kept.
ATCC Medium 0802
Cerophyll*........................................................2.5 g
Distilled water..................................................1.0 L
Combine ingredients and boil for 5 minutes. Filter cerophyll solution through cheese
cloth to remove particulates. Once cerophyll solution is filtered add:
Na2HPO4……………………………………..0.5 g
Autoclave for 15 minutes at 121?C.
*Cereal Grass Media (Catalog # 9448604) from ScholAR Chemistry is similar to
Cerophyll and can be used as a substitute.