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  • Shorter forward reads than reverse

    Hello,
    I'm DanielSai from Australia.

    I did a next gen run on my sample using the illumina miseq machine. I'm getting shorter forward sequence reads than the reverse reads. What could be the problem and how can it rectified in subsequent runs?

  • #2
    You have not given much details but one reason would be sequencing configuration for example, R1 was set to less cycles than R2.

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    • #3
      There is no logical reason (other than an assymetric run setup mentioned by @nucacidhunter) to get shorter R1 reads compared to R2. What kind of sequencing are you doing and what is the run setup?

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      • #4
        Have the reads been trimmed?

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        • #5
          Originally posted by kmcarr View Post
          Have the reads been trimmed?
          Yes, reads have been trimmed

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          • #6
            Originally posted by DanielSai View Post
            Yes, reads have been trimmed
            Then it is odd that only R1 was trimmed (or trimmed to a larger extent than R2). If you have short inserts (and thus read-through into adapters on other end) then R2 reads should also be trimmed to a similar extent.

            You still need to address questions in other comments above.

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