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  • Comparing RNA levels in tumor/normal samples using RNA-seq

    Hi,

    I'm starting a new cancer gene expression project using Illumina
    RNA-seq fastq files for matched tumor/normal samples.

    The experiment is geared toward comparing the RNA levels to see
    if there is expression of some particular genes within the cancer
    samples.

    I have a few questions:

    How do I determine if there are adapter sequences within the reads?
    Will I need to trim these out?
    What is the best way to remove them?

    For efficiency, I plan to map the reads to only the particular genes
    of interest as the reference genome and not map all reads to the
    entire human genome/transcriptome using STAR aligner.

    Do I need to include a few house-keeping genes in the mapping part
    for normalization between tumor and normal and/or for normalization
    across all tumor samples?

    Are there any concerns about using the STAR aligner?

    What is the best software tool to use for comparing the various gene
    expression levels across all the samples?

    Thanks in advance for any advice.
    --
    Paul N. Hengen, PhD
    http://www.linkedin.com/in/hengen

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