Agrobacterium mediated rice transformation

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Immature embryo (12-15 days after pollination ) ---->

Precultured on ND2 medium for 4 days at 26C in darkness ---->

Immersed in the bacterial suspension for 20 minutes ---->

Incubated on ND2-As medium for 3 days at 26C in darkness ---->

Cultured on ND2CCH25 medium for 2 weeks for selection ---->

Subcultured on ND2CCH50 medium for 4 weeks ( 2 generations ) for
selection ---->

Resistant calli were put on 3M filter paper overnight for osmotic
treatment ---->

Resistant calli were transferred to NN.1B2H medium for regeneration
---->

Regenerated seedlings were transferred to MS0 medium for further growth
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Plantlets about 10cm high were grown to maturity in greenhouse in soil