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  • Sciara_coprophila
    Junior Member
    • Jun 2011
    • 1

    New to Bioinformatics following sequencing

    Hello all,

    I'm a first year graduate student and just picked/joined my thesis lab. I have a bunch of illumina DNA sequencing data for both genome assembly and ChIP-Seq-like experiments. Im completely new to the computational aspects in these approaches - e.g. I have recently taught myself to move around in a Unix environment and my only achievement so far was to run fastQC on the sequence reads. Also, I have been working toward learning python.

    What I would like to do next is align reads (ChIP-seq type experiment) to a human reference genome. Should I use MAQ? Bowtie? BWA?

    How about peak callers? PeakSeq?

    In the future I will also be dealing with RNA-seq data for both transcriptome assemblies and differential expression analyses. Any help here would also be welcome. e.g. how does one take RNA-seq data from two different tissues through the analysis pipeline and end up with those nice log-log plots?

    Many many thanks,

    Sciara C.

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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.
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  • 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...
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