Media for Lab 3.1

Submitted by kdorfman on Wed, 10/12/2011 - 15:50

Note: very small pellets, not too much green, except with IPTG. Dissapointing. Get a better pellet with 6 mL of culture + 4 minute spin (max speed = 500 with swinging bucket rotor). But still no obvious induction with lactose.

minimal medium + Kanamycin (50 µg/mL)
Aliquots in freezer are labeled in 1, 3, 6, 10 mg/mL

Adding Kanamycin to __mL medium

Kan mg/mL 10 25 50 100 250 500 1000 mL medium
1 0.5 1.25 2.5 5 12.5 25 50 uL Kan
3 0.17 0.42 0.83 1.67 4.17 8.33 16.67 uL
6 0.08 0.2 1 0.42 0.83 2.08 4.17 8.33 uL
10 0.05 0.125 0.25 0.5 1.25 2.5 5 uL

Sugar solutions

Ingredient vendor cat # MW mM 10 25 30 50 100 mL
glucose 180.15 500 0.9 2.25 2.7 4.5 9 g
galactose 180.15 500 0.9 2.25 2.7 4.5 9 g
maltose 342.3 500 1.8 4.5 5.4 9 18 g
lactose 342.3 500 1.8 4.5 5.4 9 19 g
IPTG (100mM) 0.4 0.004 0.01 0.012 0.2 0.4 mL

Each group needs

medium mL per group ~mL total
minimal 3 100
lactose 1 50
IPTG 0.75 25
gal 0.5 15
glu 1 30
malt 0.25 10

For Lab 3.2, each group needs enough as if to fill two rows of a plate with 100µL = 2.4 mL