Validating microarray data using rt real time pcr
This is not an indication of a security issue such as a virus or attack.It could be something as simple as a run away script or learning how to better use E-utilities, for more efficient work such that your work does not impact the ability of other researchers to also use our site.For each platform, four technical replicates were performed on the same total RNA samples according to each manufacturer's standard protocols.For Agilent arrays, comparative hybridization was performed using incorporation of Cy5 for brain/lung/liver RNA and Cy3 for UHR RNA (common reference).From there, we have used q RT-PCR to directly validate the gene expression using the same clinical tissue samples.
The performance of both array platforms in identifying regulated and non-regulated genes was identical.
This approach optimizes sensitivity and accuracy while controlling the cost of experiments.
A high quality c DNA array was fabricated using a restricted number of carefully selected transcripts with each clone printed in triplicate.
The current study assesses factors contributing to the correlation between these methods in five separate experiments employing two-color 60-mer oligonucleotide microarrays and q PCR using SYBR green.
Overall, significant correlation was observed between microarray and q PCR results (ρ=0.708, p), array averaging, tissue type, and tissue preparation was assessed.