SOYBEAN TRANSFORMATION PROTOCOL

(Adapted from Wayne Parrott’s protocols, http://mulch.cropsoil.uga.edu/soy-engineering/)

Induction on D40 media (6-8 weeks)

Proliferation on D 20 solid media (4 weeks)

Proliferation on FNLite liquid medium (4 weeks)

Place 20-25 clumps of SE’s on D 20 solid media

Shoot

After shooting, leave material on D 20 solid media (1 week)

Transfer material into selection media (FNLite + 20mg/L HYG) (7-10 weeks)

Isolate green tissue and inoculate into multi-well plate (5 weeks)

Remove material from multi-well plate and proliferate in FNL medium (5 weeks)

Inoculate healthy green embryos into maturation medium (SHAM) (4-5 weeks)

Desiccation 5-7 days Select healthy matured

embryos for lipid or other

Germination 1 week analysis

Plant hardening 3 weeks

Transfer hardy plantlets into pots and grow out in green house

Harvest mature seeds 10-16 weeks

See gifs of induction, transformation and conversion (& PP file)

Media comparisons