Additional file 3: of Unexpected patterns of segregation distortion at a selfish supergene in the fire ant Solenopsis invicta

Figure S1. Protocol for collecting progeny embryos for Sb supergene transmission ratio distortion (TRD) study in Solenopsis invicta. Families (progenies) of diploid embryos were obtained from individual mother queens initially isolated in plaster-bottomed petri dishes for three to four weeks with several thousand adult workers (colony fragments) (i). Queens confirmed to be mated (producing worker brood) at the end of this period were then isolated with 2–3 workers in plastic plaster-bottomed specimen cups (isolation cups) (ii). Each such queen was removed from the cup after 12 h then frozen in a − 80 °C freezer (iii). Eggs laid by the queen were maintained in the cup with the workers for an additional 48 h (by which time they were embryos within the egg coat), then transferred into a gelatin capsule and placed immediately in a − 80 °C freezer pending DNA extraction (iv). Thirty-six haphazardly selected embryos per progeny were sampled for genotyping at Gp-9 and 14 microsatellite loci (v). (TIF 1353 kb)