GeneChip® HT 3' IVT PLUS Reagent Kit

Microarray RNA amplification & Labeling - Human endometrial stromal cells Biotin

Experiment
Microarray RNA amplification & Labeling - Human endometrial stromal cells Biotin
Product
GeneChip® HT 3' IVT PLUS Reagent Kit from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
Total RNA was reverse-transcribed, amplified and labelled using a GeneChip® 3′IVT Expression Kit (Affymetrix Inc., Santa Clara, CA, USA).

Publication protocol

The RNA samples subjected to miRNA microarray analysis were also used to identify putative miRNA target genes. Briefly, total RNA from the 3 undifferentiated and 3 decidualized cultures were combined separately; and the two sets of mRNAs subjected to gene expression microarrays. Total RNA was reverse-transcribed, amplified and labelled using a GeneChip® 3′IVT Expression Kit (Affymetrix Inc., Santa Clara, CA, USA). The complementary RNA (cRNA) was purified, fragmented according to the manufacturer’s protocol and hybridised on the Affymetrix GeneChip® Human Genome U133 Plus 2.0 array. Hybridisation, washing, staining and scanning were performed using the Affymetrix GeneChip® system instruments and protocols. The expression levels of each gene obtained from the scanned data were normalised among samples using the robust multi-array average (RMA) method. The mRNAs that were differentially upregulated (>2-fold) or downregulated (<0.5-fold) upon decidualization were extracted.

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