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  • #16
    As others said, the point of ERCC is to distinguish global bias, besides technical variance. so the way you add ERCC must be based on cell number or DNA amount (equals to cell number), not RNA amount, better to add it into the lysate before RNA extraction, for less bias caused by technical issue. In that case, I don't agree with the idea spike-ins makes the data noisy.
    Spike-in should be encouraged if there is enough rationale and you do it right and carefully.
    if there is global gene expression bias, like Myc activation, then ERCC definitely give you the chance to see the difference. otherwise, just simply ignore the spike-ins, if there is no real biological bias.
    Rule No.1 again, all results must be independently reproduced before you make 'amazing' conclusion.

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