Illumina HiSeq 2500 paired end sequencing; The male determining M-factor of the global disease vector Musca domestica is a paralog of the generic splice factor CWC22
The male determining M-factor of the global disease vector Musca domestica is a paralog of the generic splice factor CWC22
All strains are described in {Dubendorfer:2002uz}{Scott:2014gi}{Schmidt:1997tw} and were cultured at 25°C in beakers filled with 140g food-mixture (150g flour, 50g yeast, 120g milk powder, 1000g bran) dissolved in 185ml water and 4ml Nipagin (66g Nipagin, 123g Nipasol in 2l of 96% Ethanol) until hatching and then transferred to cages at room temperature (RT) or kept in beakers at 18°C . Rearing of larvae and adult flies has been described in Schmidt et al, 1997. Since low density of larvae on standard medium can cause substantial decrease in survival rates, we reared injected embryos and the surviving larvae on porcine manure. To dispose of mites and other parasites and to avoid contamination with eggs or larvae from wild-type populations, manure was stored at -70° C for at least two weeks prior to use. Total RNA was prepared from 0-6 hrs and 1-8 hrs embryos with TRIzol reagent (Invitrogen) according to the manufacturer’s protocol. Libraries for sequencing were generated with the TruSeq RNA Sample Preparation kit (Illumina).