Thursday, November 13, 2003

Description
In vitro growth of seedlings

Procedure
sterilisation of seeds:

rinse with 70% EtOH for 30 sec
put in 1% bleach (sodium hypochlorite, supplemented by a few drops of Tween-20) for 5 min
wash 2-4x with sterile H2O for 1 min
plating and growth

resuspend in sterile 0.1% agarose
plate 4 ml in petridish (0.8% agarose supplemented with 1/2 MS10) in sterile hood
let dry for 1-2 h in the hood
wrap with 3M micropore tape
keep at 4C for 4 days
place the dishes under light

Recipes
Solutions
1/2 MS10

MS Macro I 50.0 ml
MS Macro II 50.0 ml
Myo 10.0 ml
FeNaEDTA 10.0 ml
MS Vitamines 1.0 ml
MS Micronutrients 0.5 ml
Sucrose 10.0 g
H2O x.x ml
Total 1000.0 ml

adjust pH to 5.7 with 1 M KOH, alternatively MES (0.5g/l) can be used to ensure more stable pH during plant growth.


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