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  • HQHong
    Junior Member
    • Oct 2018
    • 1

    in-situ (in-cell) RT

    Hi all, has anyone tried SPLiT-seq as published in "Single-cell profiling of the developing mouse brain and spinal cord with split-pool barcoding"?

    I have specific questions:
    (1) How much cells do one usually start with? I can only obtain about 10% of the cells after fixation.
    (2) How does one quality check the library? Will the amplicon size distribution be similar to conventional cDNA libraries?

    Many thanks in advance! Would be great to know if others are also trying out the same protocol

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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.
    07-01-2026, 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

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