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At the end of each probe is a wire that attaches to the skin and clothing.
One end of each probe was designed to bind to the free end of the target anthrax DNA; the other end carried a tiny gold particle.
The nonfluorescent quencher dye BHQ1 was linked to the 3′ end of each probe.
Black hole quencher (BHQ; Sigma) was attached at the 3′ end of each probe (either BHQ1 or 2, depending on the probe dye).
The front end of each probe has an aluminum flange attached (3 mm thick and 70 mm diameter), to facilitate easy fixation to the patient's chest with tape.
Figure 3B displays all probe-targeted regions in 18S and 25S rRNA (except that of the failed 25S probe), aligned according to the 3′ end of each probe, to assess depletion zone topology for different library insert size ranges.
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The reporter dye at the 5′-end of each probe was FAM.
Digestion curves suggested that trends in RNA degradation between 5' and 3' ends of each probe set were very similar.
In brief, the plasmid cDNA library was hybridized with biotinylated oligonucleotide probes (which target highly conserved regions of the α- and β-like globin genes) and blocking oligonucleotides (which correspond to the 5' and 3' ends of each probe to prevent renaturation of the plasmid DNA; Additional file 2: Table S2).
For all 631066 probes of 57381 probe sets of rice microarray, we collected their distance away from 3' end of transcript (Each probe represented by its middle nucleotide), and plotted the number of probes at the same position to show its distribution (Additional file 1: Figure S1).
The 5′ end of each oligonucleotide probe was tailed with ten dTTP molecules (T-spacer) to increase the on-chip accessibility of spotted probes to target DNA.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com