GeneChip® Human Genome U133 Plus 2.0 Array

Microarray Gene expression arrays - Human endometrial stromal cells Biotin

Experiment
Microarray Gene expression arrays - Human endometrial stromal cells Biotin
Product
GeneChip® Human Genome U133 Plus 2.0 Array from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
Avoid touching the surface of
the glass with your fingers. Skin oils and other substances, such as
lotions or ink, can fluoresce. If the surface of the glass is noticeably dirty, it can be carefully cleaned with a nonabrasive laboratory tissue.

Publication protocol

Affymetrix GeneChip™ Human Genome U133 Plus 2.0 microarrays were used for mRNA profiling. Microarray analysis was performed at Mayo Microarray Core facilities by technologists following standard procedures. Briefly, RNA samples were subject to Agilent analysis for quality controls. cDNA was prepared from 10 μg of RNA, quantified by spectrometry, and used as a template for the synthesis of biotinylated cRNA using RNA transcript labeling reagent (Affymetrix, Santa Clara, CA, USA). The quality of the cRNA probes was verified by gel electrophoresis and pilot hybridization with the Test-3 array. Hybridization solution containing fragmented cRNA probes and control cRNA (BioB, BioC, and BioD) was supplemented with herring sperm DNA and bovine serum albumin. The probe solution was heated at 99 °C for 5 min followed by incubation at 45 °C for 5 min before use. Hybridization was carried out at 45 °C for 16 h with constant rotation at 60 rpm. The arrays were washed and stained with streptavidin-phycoerythrin (Molecular Probes, Eugene, OR, USA). After washes, arrays were scanned using the GeneChip system confocal scanner (Hewlett Packard, Palo Alto, CA, USA).

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Manufacturer protocol

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