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No data?? Any advice



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  • No data?? Any advice

    Sequencing on the Illumina MiSeq cluster generation complete - but no clusters passing filter or Q30 or data generated.
    image here : https://postimg.cc/Wh3xpzCt
    I know it is over clustered but where are the (even though it is probably crappy) FastQ files? It would be nice to have a look anyway.
    The run did not terminate and completed all cycles with “no errors”.
    Any advice besides reloading the run after redoing quantification?

  • #2
    I have the same problem now, any advice/solution?


    • #3
      sequence456789 and rociogma before you loaded your libraries, did you check them on a Bioanalyzer or Tapestation? I'm curious if there tons of adapter dimers.

      What was the library prep kit used? And how did you quantify them? There's always the possibility of accidentally making a mistake during the pooling, denaturation, or loading steps.


      • #4
        I have prepared the libraries with Qiaseq kit and Qubit4, and the quality was checked by Qiaxcel Advanced.


        • #5
          rociogma that's so strange. I'm assuming the libraries looked fine on the Qiaxcel Advanced?

          One issue can be during the pooling and denaturation. I'm not suggesting that you made a mistake, but if the Tris-HCl or NaOH went bad or was at the wrong concentration then the libraries won't bind to the flow cell. If you can maybe check the pH of your NaOH and Tris-HCl.

          My main suggestion is to check the flow cell and write down the barcode number. I think you should take a picture of it for your records. I would then contact Illumina and double check there wasn't anything complaints with that batch of flow cell. Illumina always helps with stuff like this and their support staff is pretty good.


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