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  • Call of Ns in 454

    I was wondering if anyone can explain me why N would be called in the pyrosequencing, since the base detection comes in sequential flows of nuclotides.
    thanks in advance

  • #2
    An N is called when there are more than 3 flows without a signal high enough to be considered a base call. If the cycle get all the way around back to the previous base without any signal being called, then there must have been a nucleotide there that was added without the signal being strong enough to be read, so the system calls an N there because it can't know what that base was. In practical terms, N's in a read indicate a poor quality read.

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    • #3
      Thanks for you reply, that was very helpful

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