Rna clean up concentrator kit

To detect expression in the germline, abdomens were separated from the body and further dissected in PBS to extract testes or ovaries. Tissues or whole flies (5–10 total) were homogenized with TRIzol (Invitrogen) then briefly centrifuged to separate the supernatant. The supernatant was chloroform-extracted, resulting in a soluble phase that was then isopropanol-extracted. The RNA that precipitated was briefly centrifuged, and the resulting pellet was washed with 70% ethanol and resuspended in RNAse-free water. Samples were treated with DNase I (Zymo Research), followed by purification and concentration (RNA Cleanup & Concentrator kit, Zymo Research). The samples were used to generate cDNA using SuperScript III First-Strand Synthesis, following the manufacturer’s instructions (Invitrogen). The cDNA was then used to conduct RT-PCRs with Phusion (NEB). Primers are provided in Data S1.