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  • Rosanne82
    Junior Member
    • May 2009
    • 1

    3'UTR library or random primed cDNA library for quantification?

    I am quite new in the field of next generation sequencing and I hope to get some information and help here.
    I want to do a RNA-Seq by using the Illumina platform to analyze the transcript profile of a specific tissue of a wildtype plant compared to a mutant. The aim is identifying a different transcription level of some genes as well as finding some rare genes. The organism I am working with is not sequenced yet, but it should be till the end of this year.
    My question is which type of library is more suitable for quantification, a random primed cDNA library or a 3’UTR library, which proposed me a company. But what would be the advantage of a 3’UTR library?

    Thank you very much!

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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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