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  • normalizing small RNA libraries

    Hi all,
    Im currently building small RNA libraries from the same animal tissue, but after different treatments. We're trying to see if small RNAs in this tissue are differentially regulated because of the treatment. The problem im having is a normalization parameter. I thought of spiking-in constant amounts of a non- related plant or bacteriophage small RNA, into the total RNA from each treatment sample, and then making the libraries..
    Has any one done this before? Or are there better ways to normalize your samples so that one can directly compare expression levels of small RNAs?
    Appreciate your help!

  • #2
    For RT PCR ABI and Qiagen both recommend snoRNA controls for normalization

    http://www.gene-quantification.de/AB...g-controls.pdf

    http://www.qiagen.com/products/mirnacontrols.aspx

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    • #3
      Thanks for your reply. Both the ABI and the Qiagen kits use sno RNAs as endogenous controls for the QRT PCR assays. These are much longer than 40 nt but my libraries are restricted to RNAs between 15 to 30 nt. So, snoRNAs are excluded..which is why I think I need a spike in control..

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