RNA Quality Assessment
As RNA quality assessment is a critical aspect of any gene expression experiment, the quality of RNA samples delivered to the Gene Expression Core is assessed using the Agilent 2100 Bioanalyzer. This method may not be appropriate for highly degraded RNA samples, in which case other quality control metrics may be used.
By determining a priori the level of degradation within a sample, researchers can select those samples that can provide the highest quality gene expression profiling data.
mRNA. The Agilent 2100 Bioanalyzer uses microcapillary electrophoresis and lab-on-a-chip technology to separate molecules by size, providing information on the quality of a eukaryotic total RNA sample by characterizing the various RNA types. The data output includes an electropherogram that displays molecular peaks, a virtual gel image and the specific values that describe each sample.
This assay provides a numerical assessment of the quality of an RNA sample on a scale of 1 to 10. Only 1 to 1.5 microliters of total RNA is required for the analysis.
MicroRNA. The Agilent Small RNA Chip determines the percentage of small RNA (microRNA, tRNA, 5S and 5.8S rRNA, respectively) in total RNA or enriched small RNA samples. This assay requires 1 to 1.5 microliter of sample and can be used for quality assessment prior to downstream applications such as microRNA arrays and real-time quantitative PCR.
Note that highly degraded RNA may contribute to false readings due to degradation products that migrate at the same rate as small RNA.
The cost for each project is dependent on experimental design, vendor costs and time required for completion. This information is provided in each project proposal, which must be approved by the principal investigator prior to project initiation.
For more information or to initiate a project, contact us at 507-255-1086 or by email.