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What steps are necessary to construct the probe?
To construct the probe, the GNP surfaces were coated with single-stranded DNA (ssDNA) by forming Au S bonds.
This result is expected if the binding efficiencies of transcripts of the test species P. euphratica and P. × canescens with genes used to construct the probe sets on the array are similar.
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Functionalizing 13-nm GNPs with ssDNA was a crucial step in constructing the probe used in our experiment.
The sequences used to construct the Affymetrix probe set were downloaded from the GrainGenes database.
Primers were designed from the EST sequence used to construct the Affymetrix probe to produce 250 bp amplicons.
In transgenic lines expressing the δOAT-GFP fusion construct, the δOAT probe detected the native mRNA and a band with higher molecular weight corresponding to the δOAT-GFP transcript.
In order to self-consistently calculate the electrical properties of nanodevices, we construct the two-probe device geometry where the central region contains the SW defects and both leads consist each of the two supercell pristine ZGNR, as shown in Figure 1.
Apart from intact construct, the immunoblots probed with anti-OmpA antibody revealed a strong band around 17 kDa (the expected size of the periplasmic domain) that fractionates to the soluble fraction (Figure 4B).
To construct the fluorescent probes, pairs of specific primers were designed to amplify regions of the GB12449 and GB12811 DNA sequences comprising 300 and 379 nucleotides, respectively (Additional file 2: Table S2).
Thus, this work provides a promising strategy to construct the ratiometric fluorescent probe.
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