From OpenWetWare
Dropout Media
Amino Acid Dropout Powder
Mix in a dry mortar. Grind up thoroughly with a pestle for ~15 minutes. Store in 50mL conical tube.
| ADE
| 0.25g
|
| LEU
| 5g
|
| p-aminobenzoic acid
| 0.1g
|
| Add 1g each of the following:
|
| ALA
| 1g
|
| ARG
| 1g
|
| ASN
| 1g
|
| ASP
| 1g
|
| CYS
| 1g
|
| GLN
| 1g
|
| GLU
| 1g
|
| GLY
| 1g
|
| HIS
| 1g
|
| ILE
| 1g
|
| Innositol
| 1g
|
| LYS
| 1g
|
| MET
| 1g
|
| PHE
| 1g
|
| PRO
| 1g
|
| SER
| 1g
|
| THR
| 1g
|
| TRP
| 1g
|
| TYR
| 1g
|
| Uracil
| 1g
|
| VAL
| 1g
|
Dropout Liquid Media
Solution recipe for 1L liquid AA dropout media
| 1. Measure 800mL millipore water into a 2L plastic beaker.
| 800mL H2O
|
| 2. Weigh 6.7g Yeast Nitrogen Base (w/o Amino Acids) and add it to the mixture
| 6.7g Nitrogen Base
|
| 3. Weigh 2g of Amino Acid Dropout Powder and add it to the mixture
| 2g AA DO Powder
|
| 4. Stir until completely dissolved
|
|
| 5. Bring solution to 900mL with millipore water in a graduated cylinder
| Volume to 900mL
|
| 6. Autoclave 30 minutes to sterilize
|
|
| 7. Add 100mL of sterile 20% Glucose prior to use
| 2% Glucose
|
Dropout Plate Media
Solution recipe for 1L agar plate AA dropout media
| 1. Measure 20g Bacto Agar into a 2L flask.
| 2% w/v Agar
|
| 2. Pour 700mL millipore H2O into the flask containing agar
| 700mL H2O
|
| Add the following reagents to a separate beaker
|
| 3. Add 150mL millipore H2O to a separate beaker
| 150mL H2O
|
| 4. Weigh 6.7g Yeast Nitrogen Base (w/o Amino Acids) and add it to the beaker
| 6.7g Nitrogen Base
|
| 5. Weigh 2g of Amino Acid Dropout Powder and add it to the beaker
| 2g AA DO Powder
|
| 6. Bring volume in beaker to 200mL and transfer to a 500mL flask
| Volume to 200mL
|
| 7. Autoclave both flasks (nutrients & agar) 30 minutes to sterilize
|
|
| 8. Combine the 200mL nutrient flask into the 700mL agar flask aseptically
|
|
| 9. Add 100mL of sterile 20% Glucose prior to pouring
| 2% Glucose
|
