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Can anyone explain....

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  • Can anyone explain....

    Hi

    Has anyone heard of a barcoding effect on the comparability of data in the bioinformatic analysis from loading a group of samples in different lanes on a HiSeq 2500? I have heard this twice now from researchers and I am not sure what they mean...nor can I find a publication discussing this...any thoughts?

    I am very new to this world (3 months into this new job) and typically at my lab we don't offer a sequencing plan or lane mapping to researchers... we load samples that have been QC multiple time thru library construction and then they go thru qPCR and get pooled then loaded for sequencing accordingly.

    I'm trying to understand why a researcher or their bioinformatician would be worried about all of their samples not being loaded into one lane or across many lanes? Or why they would be concerned and want their samples run on the sequencer in a certain way?

    Thanks in advance for your input.

  • #2
    Pooling wrong tags can potentially lead to failure of registration of clusters during the sequencing of the tag read. Illumina has specific recommendations on pooling schemes that should be considered (this example is for nextera but you will get the idea). Tech note: http://res.illumina.com/documents/pr...guidelines.pdf

    Running a large pool across several lanes/flowcells can address potential lane/run effects as opposed to running smaller pools on specific lanes.
    Last edited by GenoMax; 11-26-2013, 08:08 AM.

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    • #3
      Originally posted by GenoMax View Post
      Pooling wrong tags can potentially lead to failure of registration of clusters during the sequencing of the tag read. Illumina has specific recommendations on pooling schemes that should be considered (this example is for nextera but you will get the idea). Tech note: http://res.illumina.com/documents/pr...guidelines.pdf

      Running a large pool across several lanes/flowcells can address potential lane/run effects as opposed to running smaller pools on specific lanes.
      Thanks for your response!

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