This Information Applies To: 2100 Bioanalyzer, 2100 Expert software, RNA 6000 Nano kit (p/n 5067-1511), RNA 6000 Pico kit (p/n 5067-1513)
Issue:
Which assay should be selected in the Agilent 2100 Expert software when running yeast total RNA samples on the Agilent 2100 Bioanalyzer system?
Resolution:
The Bioanalyzer total eukaryotic RNA assays are validated for eukaryotic RNA samples with typical 18S and 28S ribosomal RNA (rRNA) peaks. Yeast is a eukaryote but has 18S and 25S rRNA peaks. When running the total eukaryotic RNA assays, the 25S rRNA peak will migrate faster than the expected 28S rRNA molecule. Therefore, it will migrate outside the assays preset detection window for the 28S peak. As a result, the 2100 Expert software will assign the 25S rRNA fragment incorrectly, which will typically lead to unreliable RIN values or RIN, values not being displayed.
The setpoints for the prokaryotic assays are better at detecting yeast rRNA peaks. Therefore, it is best to run yeast RNA samples with one of the prokaryotic assays. Specifically, run with the Prokaryote Total RNA assay Nano Series II assay if you are using the RNA 6000 Nano kit and with the Prokaryote Total RNA assay Pico Series II assay if you are using the RNA 6000 Pico kit.
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Caution: Agilent cannot provide specifications for yeast samples as the Bioanalyzer assays were not developed for yeast RNA samples.
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Learn about RNA assays on the 2100 Bioanalyzer:
RNA Integrity Number (RIN) – Standardization of RNA Quality Control
RIN Not Calculated or NA for Total RNA Samples in the Agilent Bioanalyzer 2100 Expert Software |
For Research Use Only. Not for use in diagnostic procedures.
PR7000-8634
