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  • miRNA Sequence Read Number?

    Hello,

    I am currently setting up my first SOLiD sequencing experiment, and wondered if anyone had any information regarding the recommended number of reads per sequence.
    I am looking at ~20bp bovine microRNA and am likely to find between 250-400 unique sequences out of the RNA that I am sequencing, with a total of 10 separate samples to be sequenced at once. My options are one quarter of a plate, which I am told is equivalent to about 25 million reads, or a half of a plate for 50 million reads.
    I can't seem to find any papers that outline this process, so was wondering if anyone knew how many reads per sequence is considered necessary to have good quality data?
    Thanks!

  • #2
    In general protocol of SOLiD v3.0, the beads density should be 150K/panel.

    So you can calculate the beads number ( tags number after sequencing ) depend on which kind of slide you run.

    Full slide: 2357 panels = 354M tags (FG, *2 for MP )
    1/4 slide : 459 panels / spot = 69M tags (FG, *2 for MP )

    notics that the calculation is according to the beads density and the running quality.

    Hope it can help you.

    Comment


    • #3
      Thank you! That was the missing piece that I needed.

      Comment


      • #4
        I believe the recommended amount is 130 to 135K per panel. Be aware as well that only 50 to 60% are mappable.

        Comment

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