Chromatine immunoprecipation of atonal binding sites in Drosophila L3 eye imaginal discs. Chromatine immunoprecipation of atonal binding sites in Drosophila L3 eye imaginal discs

Atonal is a proneural transcription factor expressed in the Drosophila eye imaginal discs. To characterize the putative targes of Atonal in the eye discs we have used an endogenously GFP-tagged version of atonal to immunoprecipate disc samples with anti-GFP antibodies Overall design: L3 larvae of the Ato::GFP genotype where dissected to collect the discs. The fixed sample was homogenized by sonication. A 10% of the sonicated volume was processed as input controls while the rest was Immunoprecipitated with antiGFP antibodies, purified and sequenced.

无音调蛋白（Atonal）是一种在果蝇（Drosophila）眼成虫盘（eye imaginal discs）中表达的成神经转录因子（proneural transcription factor）。为了鉴定Atonal在眼成虫盘中的潜在靶标，我们使用了内源绿色荧光蛋白（GFP）标记的atonal基因变体，通过抗GFP抗体对眼成虫盘样本进行免疫沉淀。
整体实验设计：解剖Ato::GFP基因型的三龄幼虫（L3 larvae）以收集眼成虫盘。将固定后的样本经超声破碎进行均质化处理。取10%的超声处理体系体积作为输入对照，剩余样本经抗GFP抗体免疫沉淀后，纯化并测序。