Reagents 2012

Fisher DF0446-17-3 500 g $40.61. LB Broth Miller Luria-Bertani
25 g/L

56.25 g LB
2.25 L water

stir (takes a while)

Divvy up into 3 1 L media bottles.

autoclave 45 min (liquid cycle)

Aliquot 5 mL for students

LB agar calculators

90 plates (3 plates/pair + plates to start cultures from frozen stocks)

25 mL/plate (=~2250 mL LB)

25 g LB + 15 g agar /liter

Make 3 bottles of 750 mL each:
56.25 g LB
2.25 L water

stir (takes a while)
autoclave 45 min (liquid cycle)

put 750 mL in each of 3 1 L media bottles
add a stir bar
add 11.25 g bacto-agar
autoclave 45 min (liquid cycle)

put bottles on stir plate near sterile hood until handle-able

dispense 25 mL into sterile 100 mm Petri plates

At least two days before class: streak wild type onto LB plates

Night before class:
Pick a colony into 5 mL LB 3X
Shake at 37C

Morning of class (~10 am):
Pour each 5 mL ON culture into 125 mL LB
shake at RT
OD = ~1 when class starts
adjust heat or dilute cells to keep them from going beyond OD=1

10% NaN3 Sodium azide stock
5 g in 50 mL water

Aliquot ~750 µL per pair