Yu:Dropout Media

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(DO Media)
Current revision (19:02, 7 May 2011) (view source)
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|8. Combine the 200mL nutrient flask into the 700mL agar flask aseptically
|8. Combine the 200mL nutrient flask into the 700mL agar flask aseptically
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|Combine solutions
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|9. Add 100mL of sterile 20% Glucose prior to pouring
|9. Add 100mL of sterile 20% Glucose prior to pouring
|2% Glucose
|2% Glucose
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Current revision

Dropout Media

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Contents


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
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