Dataset features


Application: Gene expression microarray analysis
Number of samples: 12
Release date: Dec 31 2016
Last update date: May 11 2017
Access: Public
Diseases: Glioma, Neoplasms
Dataset link Characterization of Arginase Expression in Glioma-Associated Microglia and Macrophages

Experimental Protocol

The Affymetrix GeneChip Mouse Gene 1.0-ST array (Affymetrix, Santa Clara, CA) was used to define gene expression profiles from the samples. Synthesis and labelling of cDNA targets, hybridization and scanning of GeneChips were carried out by the Microarray Core Facility at the City of Hope. Due to the limited starting material, we used a modified two-cycle amplification protocol. Briefly, cRNA was generated using 10ng total RNA according to the manufacturer's protocol by using Affymetrix's GeneChip Whole Transcript Sense Target Labeling Assay in the first cycle of amplification. The cRNA was subjected to RiboMinus kit (Invitrogen, Carlsbad, CA) to remove rRNA. The resulted cRNA was used as the template for another round of amplification using the sense target labeling assay kit. Hybridization cocktails containing 5.5 g of fragmented, end-labeled cDNA were prepared and applied to GeneChip Mouse Gene 1.0 ST arrays. Hybridization was performed for 16 hours, and the arrays were washed and stained with the GeneChip Fluidics Station 450 using FS450_0007 script. Arrays were scanned at 5-m resolution using the Affymetrix GCS 3000 7G and GeneChip Operating Software v. 1.4 to produce .CEL intensity files.








Xiwei Wu

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