Pollen Tube Germination Media

0.01% boric acid
1 mM MgSO₄
2mM CaCl₂ --this was used with media containing GABA. If no other nitrogen source is present, use 1mM CaCl₂ and 1 mM CaNO₃
18% sucrose
pH 6.5

Air dry flowers for 1-2 hours (optional--this helps with release from anthers but is not essential), then dab anthers onto surface of a 25-30µL drop of germination media. Drop can be on a depression slide or on a coated slide designed to keep samples separate. Use 1 flower per drop. Invert and incubate overnight in 100% humidity. Then place small drops of vacuum grease on the slide and gently place a coverslip on top. Allow the coverslip to just touch the drops of liquid but not to squish them. Hicks et al. Plant Phys 134: 1227-39. 2004

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