Method prepared by Grace Panganiban: gepangan@facstaff.wisc.edu


Trp- B-Min Dropout Plates (2 liters)
Component A
21 g K2HPO4
9 g KH2PO4
2 g (NH4)2SO4
1 g Sodium citrate-2H2O
20 mls of 50X dropout solution (over Grace's bench)
40 mls 25X Tyrosine (over Grace's bench)
diw to 957 mls

Component B
40 agar
diw to 1 liter

After autoclaving, mix A and B, split back into 1 liter units, and add to each flask:
10 mls 100X uracil (over Grace's bench)
1 ml 20% MgSO4-7H2O (over Grace's bench)
0.5 ml 1% thiamine (in fridge on Grace's shelf)
10 mls 20% glucose (over Grace's bench)

Mix thoroughly, cool to 55-60oC, and add
1 ml 100 mgs/ml ampicillin (on -20oC freezer shelf)

50X Amino Acid Dropout Stock Solution (g/l)(can be autoclaved)
Adenine hemisulfate 1.0 Leucine 1.5
Uracil 1.0 Lysine 1.5
Inosine 1.0 Methionine 1.0
Arginine 1.0 Phenylalanine 2.5
Histidine 1.0 Threonine 10
Proline 2.5

25X Tyrosine (can be autoclaved)(can be autoclaved)
0.75 g/l