tissue: mammary gland, liver, cervix, testis, brain cell type: lipoblasts, macrophages, histocytes, lymphoblasts, B lymphocyte cell line: mammary gland adenocarcinoma, hepatoblastoma, embryonal carcinoma, cervix adenocarcinoma, brain glioblastoma, melanoma, liposarcoma, histiocytic lymphoma, lymphoblastic leukemia, plasmacytoma, myeloma disease state: control other: Cell lines derived from the various tissues and cell types
Growth protocol
n/a
Extracted molecule
total RNA
Extraction protocol
Total RNAs were extracted with RNeasy mini kit (Qiagen) according to the manufacturer’s introduction. The integrity of the RNA samples was verified using the RNA 6000 Nano Assay (Agilent Technologies, Palo Alto, CA, USA).
Label
Cy3
Label protocol
The Agilent’s low RNA input linear amplification / one-color labeling kit were used for target preparation essentially followed the protocol recommended by Agilent Technologies. Briefly, 1000 ng of total RNA was reverse transcribed into cDNA using the T7 promoter primer. Cyanine 3-dCTP labeled cRNA was synthesized from the cDNA by using the T7 RNA polymerase. To remove unincorporated nucleotides, the labeled cRNA was purified using the RNeasy mini kit (Qiagen).
Hybridization protocol
Array hybridization was performed in 100 ul of a hybridization mixture containing cRNA probes at 65℃ for 17 hrs.
Scan protocol
Slides were scanned immediately after washing on the Agilent G2565BA DNA microarray scanner using one color scan setting for 4x44k array slides (Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 100%).
Description
Gene expression of human universal reference RNA, biological replicate 2 of 2.
Data processing
Fluorescence intensities of spots were quantified, background subtracted, and dye normalized by Feature Extraction software, version 8.1.1 (Agilent Technologies). The data were then imported and analyzed using GeneSpring GX (Agilent Technologies) to generate gene lists of differentially expressed (change in expression with fold change > 2).
