This information applies to: Agilent AriaMx real-time PCR system, and AriaMx real-time PCR software
Issue
After completing an AriaMx qPCR run, no fluorescence signal is observed in either the Graphical Displays or Raw Data Plots screen.
Resolution
The absence of any signal, even a background signal, may indicate that no data collection point has been specified in the thermal profile. This issue can occur if a data collection marker is accidentally removed from one of the default thermal profiles, or if a custom profile is created without specifying data collection points. On the Thermal Profile screen, check that all required data collection markers are present.
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Caution: If a data collection marker is not added to the thermal profile, no data will be collected and the assay will need to be repeated.
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In Figure 1, fluorescence was only collected for the dissociation curve and not for the amplification segment, resulting in only dissociation curve data being available.
Figure 1. No data collection point defined in the amplification section of the thermal profile.
1. Data collection marker missing in the amplification segment, 2. Data collection marker present in the melt segment.
Figure 2 shows the same thermal profile with the correct data collection settings.
Figure 2. Data collected in both the amplification and melt sections of the thermal profile.
1. Data collection marker for the amplification segment 2. Data collection marker for the melt segment.
If the resolution in this article has not helped, contact Agilent support and provide the .amxd data file where you had this issue. To find the correct contact, go to the Agilent Contact Us page and select your country from the select a region or county drop-down list.
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Learn how to effectively operate your Agilent AriaMx real-time PCR system:
AriaMx Real-Time PCR System Setup and User Guide on Agilent.com Aria Real-Time PCR software User Manual on Agilent.com |
For Research Use Only. Not for use in diagnostic procedures.
PR7001-3588
