reagents for 3.2

Submitted by kdorfman on Tue, 10/18/2011 - 15:24

2013 - run program for 12 hours. long enough to get past stasis, and show that fluorescence continues to increase.

Give each group of 4 5 mL of each medium

Let them mix in microfuge tubes if they want.

NOTE: plate lid got stuck on PS2 - data completely worthless. Try grinding down the bottom, different lids, tape. Do a dummy run in the morning.

Had to raise up the detector by putting slices of blue ruler under its frame. Also tape the lid to the plate, and the plate to the tray.

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

If students loaded a whole row with a single solution, they would need 1.2 mL.

antibiotic calculator

Aliquot ~1.5 mL of each medium:

Totals for both days (25 groups)= 37.5 mL

Make 50 mL each
(If there are any left over from the previous year, make ~35 mL, because that is how much can be sterilized in one go with the 35 mL syringe + filter.)

Adding Kanamycin to __mL medium

Kan mg/mL 10 25 35 50 100 250 500 1000 mL medium
1 0.5 1.25 1.75 2.5 5 12.5 25 50 mL Kan
3 0.17 0.42 0.595 0.83 1.67 4.17 8.33 16.67 mL
6 0.08 0.21 0.252 0.42 0.83 2.08 4.17 8.33 mL
10 0.05 0.125 0.175 0.25 0.5 1.25 2.5 5 mL

Sugar solutions

Ingredient vendor cat # MW mM 35 50 100 250 mL
glucose 180.15 500 3.15 4.5 9 22.5 g
galactose 180.15 500 3.15 4.5 9 22.5 g
maltose 342.3 500 6.3 9 18 45 g
lactose* 342.3 500 6.3 9 18 45 g
IPTG (100mM) 0.1 0.035 0.05 0.1 .25 mL

Filter sterilize! Do not autoclave media with sugars!
* lactose takes a long time to go into solution. heat and stir overnight, covered with parafilm.