Lab 01
Media Preparation
Objective: Students will prepare media for upcoming experiments
Tobacco (hormone effects) Media
MS salts
Myo-Inositol
GM stock
Sucrose
PPM
Add per 500 mL
2.15g
50 mg
5ml
15g
0.375ml
Benzyladenine (BA) (stock)
indole acetic acid (IAA) (stock)
0.5mg |0mg |0.5mg |0mg
0mg |0.5mg |0.5mg |0mg
Adjust pH to 5.6, bring to volume in graduated cylinder 500 ml
Add 4g of agar.
Autoclave
Pour into 100x15 plates
Tobacco Transformation Co-Cultivation Media
MS salts
Myo-Inositol
Sucrose
Thiamine stock
IAA stock
Kinetin stock
Add per 500 mL
2.15g
50 mg
15g
2 mg
0.5 mg
1mg
Adjust pH to 5.7, bring to volume in graduated cylinder 500 mL
Add 4g of agar.
Autoclave
Pour into 100x15 plates
Selection antibiotics (added after media has been autoclaved and cool to the touch)
Claforan (stock)
Kanamycin (stock)
200mg/L
50mg/L
Attention: Be sure to use an overflow pan when autoclaving!
For pouring Petri dishes: mix well on stirring plate. Cover beaker with double aluminum foil,
label and autoclave at 121C for 30 minutes. Place beaker on stir plate under hood, loosen foil
cap, cool slightly and pour petri dishes. Before pouring, add Antibiotics (if applicable) to media
when cool to touch. Observe sterile technique.
GM stock (per 1 liter)
Glycine
40mg
Nicotinic Acid
100mg
Pyridoxine HCl
Thiamine HCl
200mg
200mg