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  • ilivyatan
    Junior Member
    • Aug 2010
    • 7

    Splice aware RNA-seq analysis

    Hi,
    I have RNA-seq data generated on ABI's SOLiD platform (i.e. colorspace) and have analyzed the basics using the bowtie aligner. Now I want to use a splice-aware mapper to map reads to exon junctions.
    I noticed that Tophat doesn't yet support SOLiD data.
    Does anyone have a suggestion ab out how to go about this?
    It seems that there are some bugs when using Bioscope output for Cufflinks - has this worked for anyone?
    Should I convert my data to fastq and work from there or am I going to lose too much accuracy?

    Any help would be appreciated!
    Thanks,
    Ilana

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  • GATTACAT
    Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
    by GATTACAT
    Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
    Yesterday, 11:43 AM
  • SEQadmin2
    Nine Things a Sample Prep Scientist Thinks About Before Sequencing
    by SEQadmin2


    I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

    Here are nine questions we think about, in roughly the order they matter, before...
    06-18-2026, 07:11 AM
  • SEQadmin2
    From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
    by SEQadmin2


    Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


    The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
    ...
    06-02-2026, 10:05 AM

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