Error -2,147,213,486

I am in the process of running multiple high sensitivity DNA chips back to back on our 2100 Bioanalyzer and after running two chips I began receiving the following error message on almost all of the samples in each chip. I ran the suggested daily cleaning protocol from the manual and still received the same error message on more than half of the samples on the next chip. The samples are recently made (<1 week old) whole-genome shotgun sequencing libraries that we are trying to check before sending for sequencing. The reagents we are using are from a brand new reagent kit and all of our pipets were calibrated less than 2 weeks ago. Our bioanalyzer was last serviced in 2012 but has been working fine until this. Any help is appreciated.

Error Code

Error Message

-2,147,213,486

The lower marker could not be found automatically as there are no valid peaks in the detection area between 38s and 48.5s. If possible, you should try to lower the integrator height threshold or add a peak using manual integration to solve this problem.

-2,147,213,486

The upper marker could not be found automatically as there are no valid peaks in the detection area between 98s and 129s. If possible, you should try to lower the integrator height threshold or add a peak using manual integration to solve this problem.

  • Hello,

    Thank you for your post. These errors are due to the software not being able to call the Lower Marker (LM) and Upper Marker (UM) peaks correctly. There could be a number of root causes and without seeing your data, I do not want to speculate on what the root cause may be. Please share the raw data files in the .xad format with your local support team. Please visit Contact Us | Agilent to find the contact information of your local support team.  

    Best Regards,

    Ian

  • Hello,

    Follow up to our earlier response, I wanted to share that default location for .xad files is C:>Program Files (x86)>Agilent>2100Bioanalyzer>2100 expert>Data. For US and Canada, data files can be emailed to electrophoresis@agilent.com for detailed analysis.

    Also, typically, this sort of issue is not instrument base unit/hardware related but application related. Specifically, this issue is commonly resolved by addressing the following points:

    • Perform routine maintenance.

    If not done recently, please perform maintenance of the priming station following steps as outlined starting page 134 of the Maintenance and Troubleshooting (M&T) guide and as described in our Agilent Knowledge Portal article here . Important: Please ensure the chip priming station settings are set according to the kit guide including the base plate position and syringe clip.

    Please also perform thorough cleaning of the pin set as described on page 128 of the M&T guide above and execute the short circuit test after ensuring that the pin-set is clean and dry. Additionally, here is a video that describes the process.

    • Please ensure that samples are loaded within the listed/validated specifications for the kit in use including but not limited to sample sizing/concentration range and compatible level of salt in sample buffer. In this case, please refer to DNA High sensitivity kit guide.

    We are happy to comment further once we receive your raw data example files.

    Thank you, Shweta

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