ATCC medium: 1011 Diphasic Blood Agar Medium
Complete Medium
Blood Agar base (ATCC Medium 449)……………………………700 ml
Rabbit Blood, defibrinated………………………………………….300 ml
Locke’s solution (added as an overlay; see below)…………..…450 ml
Cool ATCC Medium 449 (see below) until it may be held comfortably in the hand.
Aseptically add 30% sterile, defibrinated rabbit blood; dispense 5 ml per 16 X 125 mm screw-capped test tube, slant and cool; add 3 ml sterile Locke's Solution overlay (see below).
ATCC Medium 449
(composition per 750 ml)
Meat extract (dry, granulated)……………………....... 25.0 g
DI Water.......................................................................250.0 ml
Infuse beef in water by bringing to a rapid boil for 2-3 minutes, stirring constantly. Filter through Whatman #2V filter paper if required and add:
Neopeptone (BD 211681)...........................................10.0 g
NaCl.............................................................................2.5 g
Agar.............................................................................10.0 g
DI Water to...................................................................500.0 ml
Adjust pH to 7.2-7.4. Autoclave at 121?C.
Locke's Solution:
NaCl.........................................................................8.0 g
KCl............................................................................0.2 g
CaCl2........................................................................0.2 g
KH2PO4 ...................................................................0.3 g
Glucose.....................................................................2.5 g
DI Water…….............................................................1000 ml