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This microarray was hybridized with cyanine-dye labelled cRNA probes synthesized from RNA from female or male adults of H. contortus.
Depending on the experiment, each microarray was hybridized with one labeled aRNA sample or two samples labelled with different dyes.
To this end the microarray was hybridized with RNA extracted from animals that had been treated under different regimes of both cold and salt-induced dehydration.
A direct design was used for hybridization of the two-color arrays (Agilent and home-spotted), i.e. each microarray was hybridized with two RNA samples from different groups.
To assess the ubiquity and specificity of the oligonucleotide probes, the DNA microarray was hybridized in triplicate with genomic DNA isolated from either L. infantum or L. major.
Each microarray was hybridized with 20 μg of fragmented cRNA followed by incubation for 16 h at 45˚C with eukaryotic hybridization controls and herring sperm DNA.
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Microarray were hybridized with 33P-labelled probes, first with an oligonucleotide sequence common to all spotted PCR products (5'TcalledGGAAACAGCTATGAC3') called vector probe to measure the amount of PCR products spotted onto the microarray.
RNA extracted from the hypothalamus microarrays was hybridized with labeled cRNA as described above.
For mRNA expression experiments, microarrays were hybridized using the MAUI hybridization system (BioMicro), which promotes active mixing during hybridization.
DNA microarrays were hybridized in an ArrayBooster hybridization station (Sunergia Group) for 14 h at 42 °C.
Microarrays were hybridized in Agilent microarray hybridization chambers for 17 h at 60 °C with mixing on an Agilent rotator in a Robbin's Scientific hybridization oven.
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