Media for Ceratopteris transformation

Media for Ceratopteris transformation

Benomyl (BEN) 1000x stock = 3mg/ml DMSO (do not filter sterilize); 1x = 3 microg/ml. DMSO is stored in the hood

BAP 1000x stock = 1mg/ml dd water; dissolve and filter sterilize; 1x = 1 microg/ml

Kinetin (KIN) 1000x stock = 1mg/ml dd water; dissolve and filter sterilize; 1x = 1 microg/ml

Hygromycin (HYG) 1000x stock = 50 mg/ml dd water; dissolve and filter sterilize; 1x=50microg/ml

MSO (for all media) (per liter)

4.16 g MS salts

0.5g MES

20 g sucrose

pH 5.8

0.7% agar (7g)

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1)    Callus inducing medium

MSO + BAP + BEN

2)    Pre-bombardment medium (the only media to be pour into small petri dishes)

MSO + KIN + BEN

3)    Post-bombardment medium

MSO + KIN + BEN + HYG

4)    Regeneration medium

MSO + BEN + HYG