Notch targets in KC Drosophila cells

To identify genes upregulated in response to Notch signalling in KC cells. Keywords: Expression analysis at a single timepoint (30' after Notch activation) KC cells were obtained from Dr Martin Zeidler. Control versus Notch activated KC cells: mRNA was extracted from KC cells incubated in the absence or presence of EDTA to activate Notch (cells were harvested 30 minutes after addition of EDTA). RNA was isolated using TRIzol (Sigma) and reverse transcription was performed with Superscript III Reverse Transcriptase (Invitrogen) and oligo-dT primers (Sigma). Control and experimental samples were labeled with Cy3 or Cy5, mixed together and hybridized on Drosophila transcriptome long-oligonucleotide microarrays (FlyChip, FL002; http://www.flychip.org.uk/services/core/FL002/). Three biological replicates and dye swaps were performed (5 arrays in total/experiment). Slides were scanned by Genepix 400B dual laser scanner (Axon) and spots were found and quantified by Dapple software.

本数据集旨在鉴定KC细胞中响应Notch信号通路而上调的基因。
关键词：单时间点表达分析（Notch激活后30分钟）
KC细胞由Martin Zeidler博士惠赠。
实验组与对照组KC细胞的构建方案如下：将KC细胞分别于不含EDTA（对照组）和添加EDTA以激活Notch信号（实验组）的培养基中孵育，于添加EDTA 30分钟后收集细胞，随后提取mRNA。总RNA采用TRIzol试剂（Sigma公司）提取，以Superscript III反转录酶（Invitrogen公司）与寡聚dT引物（Sigma公司）完成反转录反应。对照组与实验组样本分别用Cy3或Cy5荧光染料标记，混合后在果蝇转录组长寡核苷酸微阵列（FlyChip，FL002；http://www.flychip.org.uk/services/core/FL002/）上进行杂交。实验共设置3次生物学重复，并开展染料交换实验，每个实验组总计使用5张微阵列芯片。芯片扫描采用Genepix 400B双激光扫描仪（Axon公司）完成，芯片点信号的识别与定量由Dapple软件实现。