Illumina HiSeq 2000 paired end sequencing of SAMD00089356
Stage 10-15 embryos [4 h 20 min - 13 h 20 min after egg laying (AEL)] derived from nos-Gal4 / nos-Gal4 females mated with UASp-Flag-EGFP-Sxl / UASp-Flag-EGFP-Sxl males were homogenized in ice-cold extraction buffer [EB: PBS containing 0.5% TritonX-100, cOmplete Protease Inhibitor Cocktail (Roche), and 100 U/ml Protector RNase inhibitor (Roche)] and centrifuged at 20000 x G for 10 min. The supernatant was collected and incubated with anti-GFP antibody in the presence of protein A-Sepharose beads (GE Healthcare). After incubation, RNAs were extracted by using ISOGEN (Nippon Gene) from the supernatant. Amplified cDNAs were prepared by using Ovation RNA-Seq System V2 kit (NuGEN), libraries created by TruSeq DNA v2 reagents.