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Before performing the microarray assay, the quality of the DNA was examined on a NanoDrop spectrophotometer (Thermo, Wilmington, USA).
For the microarray assay, the yeast cells were grown for 9 hours (mid-exponential phase) in 50mL of liquid YPD at 30°C with mechanical shaking (200rpm).
For an mRNA sample of 0.2 μg used in each microarray assay, the total amount of rare, intermediate and abundant species are 0.13 μg, 0.07 μg and 4.8 × 10-3 μg respectively.
Based on the results of microarray assay, the expression profile of genes involved in Rho/ROCK pathway and AKT pathway were further evaluated with quantitative real-time RT-PCR or western blot methods.
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Recognizing that nanoparticle assays have many of the same challenges as microarray assays, the Nano WG looked to the microarray community for lessons learned.
To compare qPCR-array and microarray assays, the log2 of microarray signals was used.
For example, both lncRNA and mRNA of ADAMTS15 were down-regulated in microarray assays; however, the lncRNA was down- but the mRNA was up-regulated in RT-qPCR.
This time series experiment was repeated with the wild type and a soxR deletion strain, resulting in 18 microarray assays from the three time course experiments (two sets for wild type and one set for ΔsoxR).
Probing microarray assays in the presence of a hybridization mix retrieves precious information on hybridization kinetics.
In additional HRneg/Tneg microarray assayed cohorts, the five-gene ICS also proved prognostic irrespective of primary tumor nodal status and adjuvant chemotherapy intervention.
We also estimated the false positive rate of the protein microarray assays based on the EMSA results.
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