Method for determining mating types of Dictyostelium discoideum

Contributed by Jakob Franke, December 2005
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Add 0.5 ml LP agar to each well of a 24-well plate. Allow to solidify.

Add 0.5 ml SS to each well.

Add 5 µl of an overnight culture of K. aerogenes to each well.

Add 1 sorocarp of each strain being tested to an individual well.

Wrap in aluminum foil and incubate at least 3 days at 22°C without shaking.

Some times it takes 5 days to get convincing macrocysts.

If in doubt, add 25 µl of a 10% TritonX100 stock solution to lyse the remaining cells and remove cell aggregates.

If you want to know whether the strain is NC4-like (matA1) or V12-like (matA2), test versus IR1 and IR3. If macrocysts form with IR1 (matA2) the strain is NC4-like; if macrocysts form with IR3 (matA1) it is V12-like. Bisexual strains, which we propose to call matA3, form macrocysts with both IR1 and IR3. "Selfers" form macrocysts by themselves.

References:

Clark et al., 1973 BBRC 52:672-678.
Erdos et al., 1973 PNAS 70:1828-1830.
Wallace and Raper, 1979 J. Gen. Microbiol. 113:327-337.
Francis and Eisenberg, 1993 Mol. Ecol. 2:385-391.

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