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题目:: RNAi profiling by array of human U202 cells with Cdh1shRNA knockdown to identify candidate interacting or target proteins of the anaphase-promoting complex_cyclosome
ID:
状态:: 发布时间Sept. 10, 2010 , 更新时间 June 3, 2014 , 提交时间 Aug. 11, 2010,
物种:: Homo sapiens
摘要:: U2OS cells were transduced by a lentiviral vector carrying a Cdh1 shRNA and a puromycin resistance gene for selection. A vector carrying a GFP shRNA was used as control. The cells were stored in liquid N2 within 10 days after transduction in order to minimize the number of passages. Cells were thawed one week before the experiment, and grown in up to seven 175T flasks containing 6x106 cells each. For synchronization, 2.5mM thymidine was added to the medium for 21 hours. After removal of thymidine, cells were cultured with 100nM nocodazole for 18 hours. Cells that had entered mitosis were collected, washed in PBS for 3 times, and cultured in fresh media for another 6 hours. RNA was isolated and analysed by Gene expression analysis using the HG-U133plus 2.0 GeneChip array (Affymetrix, Santa Clara, CA, USA) according to the standard Affymetrix protocol using 5µg of total RNA
实验种类:: transcription profiling by array
样本量:: 2
实验设计:
: 无设计数据
数据号:: E-MEXP-2860
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