10x DO buffer from 20x -UTHL Master Mix
*this is a new system, refer to Aaron for questions
the goal is to simplify the amount of mixing required for homemade dropout media
1. For any mixes which dropout one of UTHL, 10x DO mixes you can use the 20X stock in the fridge room.
2. Example for 1L of 10x -U DO
the goal is to simplify the amount of mixing required for homemade dropout media
1. For any mixes which dropout one of UTHL, 10x DO mixes you can use the 20X stock in the fridge room.
2. Example for 1L of 10x -U DO
- pour 500mL of the 20X stock
- add the appropriate amounts of His, Leu and Trp listed below (for 1L this is 20mL His, 20mL Tryp, and 60mL Leu)
- add DI water to 1L
- filter sterilize the mix and store in 4C room
10X DO Buffer (mixing from scratch)
Amino Acid
Adenine sulfate
Uracil - - - - - - - - - - - - - - - Tryptophan Histidine hydrochloride - Arginine hydrochloride Methionine - - - - - - - - - Tyrosine Leucine - - - - - - - - - - - - - - Isoleucine L-Lysine hydrochloride - - Phenylalanine Glutamic acid - - - - - - - - L-Aspartic acid L-Valine - - - - - - - - - - - - - Threonine Serine - - - - - - - - - - - - - - Nanopure Water |
Stock Concentration(g/L)
2
2 - - - - - - - - - - - - - - - - - - 10 10 - - - - - - - - - - - - - - - - - 10 10 - - - - - - - - - - - - - - - - - 2 10 - - - - - - - - - - - - - - - - - 10 10 - - - - - - - - - - - - - - - - - 10 10 - - - - - - - - - - - - - - - - - 10 30 - - - - - - - - - - - - - - - - - 40 80 - - - - - - - - - - - - - - - - - |
Volume Added(mL/L)
100
100 - - - - - - - - - - - - - - - - - 20 20 - - - - - - - - - - - - - - - - - 20 20 - - - - - - - - - - - - - - - - - 150 60 - - - - - - - - - - - - - - - - - 30 30 - - - - - - - - - - - - - - - - - - 50 100 - - - - - - - - - - - - - - - - - 100 50 - - - - - - - - - - - - - - - - - - 50 50 - - - - - - - - - - - - - - - - - - Remainder |
Final Media Concentration
(mg/L) 20
20 20 20 20 20 30 60 30 30 50 100 100 150 200 400 - |
Notes
-
- - - - - add ~5mL 5N NaOH - - - - Stored at RT Stored at RT - - - - |
Notes
- Leave out whichever amino acid(s) you're dropping out, and replace the volume with water.
- Stocks which are stored at RT are in the cabinet with autoclaved media.