ChIP-Seq for dRETL1 testes rescued with dRTEL1-GFP
Newly eclosed Drosophila males with dRTEL1-GFP expression were selected and aged for 3 days at 25°C. 800 pairs of testes were collected for each ChIP-seq reaction as one replicate. Two replicates were generated for control or experiment group. The whole ChIP process was conducted at 4°C or on ice. ChIP-IT high sensitivity kit (Active motif) was used to perform the ChIP experiments according to the manufacturer's instruction. The chromatin was immune-precipitated by a ChIP-grade GFP antibody (ab290, Abcam). Sonication was conducted on Bioruptor sonicator (diagenode) for 30 second on-off pulse, 30 cycles at 4°C. The fragment size of sonicated chromatin was confirmed between 200bp to 500bp. Illumina TruSeq ChIP Sample Preparation Kit (IP-202-1012) was used to generate the ChIP-seq libraries following the manufacturer's instruction. The Illumina compatible libraries were sequenced with Mi-seq desktop sequencer (Mi-Seq, Illumina) by BGI, China. 75 bp single-end read sequencing was accomplished. Sequencing reads were mapped to Drosophila genome (dm6, FlyBase 6.05).