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  • Multiplex strategies. HELP

    Hi all

    I plan to use the next gen sequencing services of an industrial company to quantify differential expression of microRNA in rat model of demyelination.
    we do have 16 individual samples from 4 different conditions (4 samples by condition) we will like to decrease the cost and use the multiplex bar code technique.
    My understanding is that we can create 16 different microRNA library and pool them afterward in 4 lanes of sequencing. (each lane by condition) and then demultiplex the data for analysis.
    However since the generation of the library is as costly as the sequencing, can we have a different approach and create 4 libraries (each library representing the pooled sample -4- of each condition, and each of the 4 sample in a single library being attached with a bar code) and then run 4 lanes of sequencing?
    this will save is almost 4000 dollars but i'm unsure about the lost in quality and the accuracy of the data afterward.

    thanks for helping a newbie in the nexgen sequencing world.

  • #2
    so will this will be cDNA resequencing? I am unsure from your description.

    you could try scaling down (1/2 or 1/4) reaction volumes for library preparation; however, i don't have personal experience with this.

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    • #3
      No this is for miRNA sequencing

      Comment

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