Yu:Dropout Media

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

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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

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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	Combine solutions
9. Add 100mL of sterile 20% Glucose prior to pouring	2% Glucose