Triple dropout formulation of Synthetic Complete Supplement Mixture (SC) of amino acids for S. cerevisiae growth media. Also known as Hopkins Mixture and is available in complete and dropout formulations.
Preparation Method
Reagents are tested and chosen for their ability to promote optimum growth of yeast strains when combined in the Drop Out Base (DOB) or Drop Out Base with 17 g Agar (DOBA) formula, with SC-His-Leu-Trp. DOB contains 1.7g Yeast Nitrogen Base, 5g Ammonium Sulfate, and 20g Dextrose per liter. Chosen reagents are combined in the proper proportions according to the formulation listed on the Certificate of Analysis and milled to thoroughly blend the reagents for a uniform distribution. A sample of the milled and blended formulation was used to prepare liquid (or plate) medium by adding 2.7g of DOB (or 4.37g DOBA) powder and 0.166 g of SC-His-Leu-Trp to a 100mL of purified water. The combinations were mixed by hand for about 1 minute to dissolve the dextrose and autoclaved at 121°C for 15 minutes. After cooling to 50°C, plates of DOBA with SC-His-Leu-Trp were poured. DOB (or DOBA) with SC-His-Leu-Trp liquid solution was visually inspected after autoclaving for complete dissolution of components. The pH of the medium was tested to insure that the proper specification was attained. Acceptable pH range is 4.0 - 4.8 for DOB with SC-His-Leu-Trp, report result for DOBA with SC-His-Leu-Trp. DOB (or DOBA) with SC-His-Leu-Trp was tested by S.cerevisiae cell growth at 30°C for approximately 48 hours.
Recommended Use
Add 1.66 g of SC-His-Leu-Trp to each liter of DOB (or DOBA) and autoclave at 121°C for 15 minutes for complete synthetic defined medium for S.cerevisiae.