ATCC Medium: 1651 MA Medium
Complete Medium
Proteose Peptone……………………………………………..10.0 g
Trypticase Peptone…………………………………………...10.0 g
Ribonucleic Acid yeast Solution (See Below)……………...100.0 ml
Alpha-lecithin solution (See Below)………………………….10.0 ml
Vitamin Solution, add after autoclaving (See Below)………2.0 ml
DI Water………………………………………………………..390.0 ml
Artificial Sea Water……………………………………………500.0 ml
Combine all except vitamin solution and autoclave at 121oC.
Once medium has cooled, aseptically add vitamin solution.
Aseptically pH complete medium to 7.2.
Ribonucleic Acid yeast Solution
Ribonucleic Acid (Sigma R6625)……………..……………...1.0 g
DI Water…………………………………………………………100.0 ml
Bring solution to boil while mixing to dissolve ribonucleic acid. Add to medium
above.
Alpha-lecithin Solution
Phosphatidylcholine (Sigma P-5394)………………………….0.2 g
Ethanol……………………………………………………………10.0 ml
Dissolve phosphatidylcholine in ethanol and slowly add to medium above.
*phosphatidylcholine will be completely clear in the ethanol but will become
cloudy once added to the medium. This is normal; according to Sigma’s product
sheet for P-5394, phosphatidylcholine will be clear to hazy once dissolved.
Vitamin Solution
Biotin, 1 ug/ml ethanol………………………………………….10.0 ml
Folic Acid…………………………………………………………50.0 mg
Nicotinamide……………………………………………………..50.0 mg
Ca-Pantothenate……………………………………………….100.0 mg
Pyridoxal-HCl……………………………………………………50.0 mg
Riboflavin………………………………………………………...50.0 mg
Thiamine HCl……………………………………………………150.0 mg
Thioctic (Lipoic) Acid…………………………………………...1.0 mg
DI Water…………………………………………………………100 ml
Stir while adding components. For long term storage, aliquot and freeze.