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  • one third of maximum florescence intensity

    Good day all,
    My question pertains to calculating one third of maximum florescence intensity (MFI)
    If the MFI is at 15 cycles then one third is 5 cycles.
    Why the need to read of the amplification plot in the linear scale.

  • #2
    I am not sure I understand your point but I am curious

    Is it at the library prep step, using PCR? If yes, your material will (almost) double at each cycle, so your asumption is wrong. If it is something else, I would be glad to learn something new today

    Comment


    • #3
      ATAC-seq

      Hi SylvainL,
      Thanks for your comment I will fill you in…

      It is not my assumption it is a quote from Curr. Protoc. Mol. Biol. it pertains to ATAC-seq.

      ATAC-seq: A Method for Assaying Chromatin AccessibilityGenome-Wide. Curr. Protoc. Mol. Biol. 109:21.29.1-21.29.9.

      “17. To calculate the additional number of cycles needed, plot linear Rn versus cycle and determine the cycle number that corresponds to one-third of the maximum fluorescent intensity (see Fig. 21.29.2A).”

      I was wondering if anyone has performed this calculation comparing linear to logarithmic amplification plots, and whether a formula exist to precisely determine “one-third of the maximum fluorescent intensity” without visually guessing from the amplification plot.

      Cheers,

      Comment

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