Reagents for 5.1

Reagents for 5.1 kdorfman Fri, 11/04/2011 - 14:49

Minimal Medium + HMLTT

Check stocks in refrigerator before making any solutions!

200x HMLTT

1 g histidine
1 g methionine
1 g leucine
1 g threonine
0.2 g thiamine

in ~80 mL H2O.

Cover with parafilm and stir for 2 hours

Bring to final volume 100 mL

Filter sterilize

10x M9 salts

112.2 g M9 salts (Difco 248510 Fisher DF0485-17 500 g $75.20)
1 L H20
autoclave 45 min

M9 Minimal Medium (glycerol) for chemotaxis strains

Aliquot ~5 mL in case they need to dilute their cultures. Make about 24 tubes, and see if it lasts through 5.1 and 5.2.

50 mL for student cultures
6 mL for overnight cultures
50 mL for dilutions, etc.
50 mL with added aspartate (0.01M)
50 mL with serine (0.01M)
50 mL with d-aspartate (0.01M)

Aspartate (or D-Aspartate) 0.01M

hard to get into solution - stir for several minutes

filter sterilize

Make 50 aliquots (~0.5 mL) to last through 5.1 and 5.2

serine 0.01M

hard to get into solution - stir for several minutes

filter sterilize

Make 50 aliquots (~0.5 mL) to last through 5.1 and 5.2

Plus, for extra-mile experiments:

0.01 M glutamic acid
pH 5.06 (adjust with acetic acid)
0.01 M galactose
1mM CoCl2

To make	mL min med
start with	mL 10X M9
add:	mL 200X HMLTT
add:	100% glycerol
add:	mL H2O
add:	mL 1M MgSO4
add:	uL 1M CaCl2, stir if it precipitates

MW of aspartate:
Concentration:	M
to make:	mL aspartate in min med
add:	g aspartate

MW of serine:
Concentration:	M
to make:	mL serine in min med
add:	g serine