The following protocols were used in the G1E Microarray Project. For the most part, these are adaptations of the methods supplied by Affymetrix. Use of particular brands does not imply that others would not work. The author has no commercial interest in these products.
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G1E-ER4 Cells: Medium,
Propagation, Estradiol Treatment, Freezing
- Preparation of RNA
- RNA Solutions
- Northern Blot Procedure
- Preparation of Probes for Northerns
- Purification of DNA from Agarose Gels
- Labeling with Roche High Prime Kit
- Spin-column Cleanup of Probe
- Sample Preparation for Gene Array Analysis
- Harvest Log Phase Cells
- Recovery of mRNACheck RNA quantity/quality
- RNA purification
- cDNA: First Strand Synthesis
- cDNA: Second Strand Synthesis
- cDNA Clean Up
- IVT of Biotinylated cRNA
- Purification of IVT Transcripts
- Fragmentation of Biotinylated Target RNAs
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