Media and solution preparation for this protocol

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== Media and solutions for Arabidopsis transformation ==

YM Media 1L Mannitol 10g Yeast extract 0.4g K2HPO4 (10% w/v stock) 1 ml KH2PO4 (10% w/v stock) 4 ml NaCl (10% w/v stock) 1 ml MgSO4.7H2O (10% w/v stock) 2 ml

• pH 6.8 • Agar 15g/L • Autoclave • When ready to pour add: • Antibiotic selection if required

Keep poured plates for 2 days at room temperature to visualise any contamination, then store at 4°C.

Minimal A Medium (Svab, et al, 1975)

Liquid 1L Stock solution A 490mL Stock solution C 11mL Milli-Q H2O 499mL

Solid 1L Stock solution A 490mL Stock solution B 499mL Stock solution C 11mL

• Store 4°C

Minimal A Stock Solution A

 Final concentration 
   g/490mL    1 x mM 

K2HPO4 10.5 60 KH2PO4 4.5 33 (NH4)2SO4 1.0 7.6 tri-Sodium citrate.2H20 0.5 1.7

• Autoclave in 490mL aliquots • Store 4°C • Substituting chemicals • K2HPO4.3H2O 13.8g/490mL

Minimal A Stock Solution B

Dissolve 15g agar in Milli-Q H2O, make up to 500mL

• Autoclave • Store room temperature

Minimal A Stock Solution C

Add 1mL 1M MgSO4 to 10mL 40% sucrose

• Store room temperature

Infiltration media

   200ml  Final conc. 

10X MS salts 20ml 1xMS salts Sucrose 10g 5% MES 0.2g 0.1% Silwet-L77 200