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The isolated RNA sample was used both for the microarray and for validation assays by qRT-PCR.
For validation assays of the identified links in the network we used a second set of 16 healthy individuals.
A dose-response was performed on all tested compounds, both for the primary screen and for validation assays.
To further illustrate this issue, we plotted the viability data against the (tend/tzero) ratios for the constructs we used for validation assays.
Target prediction algorithms are undoubtedly valuable tools to researchers in the field providing a rapid appreciation of the potential processes impacted by a particular miRNA and prioritising potential direct targets for validation assays.
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All compounds were tested as per the treatment regimen established for Validation assay I.
Finally, one gene belonging to the group of genes related with ribosomal proteins, RPS24, was selected as the housekeeping gene for the validation assays in this study.
Genomic DNA from 85 of the discovery panel fish and 98 samples from five NCCCWA mapping families [ 21] were used for the validation assays.
For the validation assays with MTS and QRT-PCR, "Yes" indicates that knockdown of the gene altered drug cytotoxicity, while "No" indicates no alteration.
Sanger sequencing operates at low throughput and also requires human interpretation of the sequencing results, making it only suitable for small validation assays.
From 28 candidate shRNAs identified from the pooled negative selection screen to potentially inhibit the viability of the breast carcinoma cell line MDA-MB-231, a subset was selected for arrayed validation assays.
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