Microarray Hybridization Protocols

Protocol describes the direct detection of RNA on DNA microarrays using Hybrid Capture (HC) technology and the HC ExpressArray Kit developed by Diagene.  The kit uses a proprietary antibody that binds specifically to RNA:DNA hybrids and a second, fluorescently labeled, antibody that detects the primary antibody.  Total RNA is applied directly to a glass-spotted DNA microarray, and stable RNA:DNA hybrids are visualized via a Cy3-labeled secondary antibody. - [Read Hybridization and Detection Using the HC ExpressArray Kit Protocol]

Protocol for hybridization to a scanning array made on glass. - [Read Hybridization to a Scanning Array Made on Glass Protocol]

Protocol details the preparation of biotin-labeled target samples and hybridization of these samples to an Affymetrix in situ synthesized oligonucleotide GeneChip array.  The procedure requires a minimum of 5 µg of purified total RNA as starting material. - [Read Microarray Protocol for Affymetrix In Situ Synthesized Oligo Arrays]

Protocol details the preparation of fluorescently labeled target samples and hybridization of these samples to a microarray of Agilent inkjet-deposited presynthesized oligonucleotides.  The procedure requires a minimum of 3 µg of purified total RNA as starting material. Includes: cDNA Synthesis; Fluorescent cRNA Synthesis; cRNA Precipitation and Cleanup; cRNA Quantification; Hybridization; Washing. - [Read Microarray Protocol for Agilent Inkjet-Deposited Presynthesized Oligo Arrays]

Protocol for the preparation of fluorescent DNA probe from HUMAN mRNA or total RNA using direct incorporation. Includes: Preparing fluoresenctly labeled cDNA (probe); Washing and Scanning Arrays. - [Read Preparation of Fluorescent DNA Probe from HUMAN mRNA/ Total RNA using Direct Incorporation Protocol]

Protocol for spotted microarray hybridization and scan. Includes: Setup Hybridization; Wash Slides; Scan Slides. - [Read Spotted Microarray Hybridization and Scan Protocol]

Protocol details the preparation of fluorescently labeled target samples and hybridization of these samples to a microarray of Agilent inkjet-deposited cDNAs.  The procedure requires a minimum of 5 mg of purified total RNA as starting material. Includes: First Strand cDNA Synthesis; Second Strand cDNA Synthesis; cDNA Cleanup and Precipitation; In vitro Transcription; Cleanup and Quantification of in vitro Transcribed RNA; Fluorescent Labeling of the Target Samples. - [Read Transcript Profiling by Microarray Analysis Protocol.]