FIGURE 5-2. Description of the MAGIChip DNA oligonucleotide microarray technology used in the Stahl laboratory.

FIGURE 5-2Description of the MAGIChip DNA oligonucleotide microarray technology used in the Stahl laboratory

(A) Simplified schematic overview of the protocol used to fragment and end-label total RNA directly from saliva. The RNA pool is complex, and targets can range in size from 100 bases to several kilobases. Fragmentation is a chemical nuclease reaction, where the average desired size of molecules ranges from 80–50 bases. (B) Image from a MAGIChip. DNA oligonucleotide microarray following hybridization with total RNA isolated from a human saliva sample. Total RNA was isolated from 300 μL of a human saliva sample. RNA was fragmented, labeled, and hybridized to the microarray. A portion of the microarray image is shown. Individual gel pads are 100 × 100 × 20 μm. Gel pads are arrayed in a 26 × 26 matrix and have 300 μm center-to-center spacing. (C) Example of nonequilibrium thermal dissociation curves measured from 3 gel pads (replicate perfectly matched probes designed to detect all life, and a probe with a SNP that is used as a reference mismatch hybridization).

SOURCE: Smoot et al. (2005).

From: 5, Addressing Complexity in Microbial and Host Communities

Cover of Ending the War Metaphor
Ending the War Metaphor: The Changing Agenda for Unraveling the Host-Microbe Relationship: Workshop Summary.
Institute of Medicine (US) Forum on Microbial Threats.
Washington (DC): National Academies Press (US); 2006.
Copyright © 2006, National Academy of Sciences.

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