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Old 06-30-2016, 08:31 AM   #1
hideandSEQ
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Location: New Haven

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Default RNAprotect Cell Reagent

So Qiagen has this RNAprotect Cell Reagent (https://www.qiagen.com/us/shop/lab-b...inginformation) which pretty much sounds like the holy grail of RNA preservation within cells.

Does anybody understand (generally) how this reagent works? Does anybody know how to pellet cells suspended in it? Particularly how to pellet cells in reagent without lysing them? If I wash out the reagent will their transcripts remain preserved? Will my expression profiles really be representative of the cell in culture? Their protocol for using it only mentions processing bulk RNA extractions via their proprietary products (which makes no sense to me because why bother to preserve transcripts within the cells if you're going to run bulk extraction anyways?)

I'm asking because I need to optimize for single cell RNA seq (Drop Seq in particular) and I've already seen some basic anecdotal tests showing it's a miracle worker for preserving transcripts in trickier cell types.

I'm also curious if anyone has any experience using this prior to FACS or with dissociated tissue suspension preparations.
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Old 07-18-2016, 04:22 AM   #2
miRnaman
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I am also interested in whether RNAProtect could work for single-cell and Drop-seq. Did you try it?
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Old 07-28-2016, 12:52 PM   #3
rnaseqmonkey
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RNAprotect Cell has been used for scRNA-seq in the SCRB-seq method (Soumillon et al 2014)
It seems to be possible to FACS "fixed" cells, but I hear this is incompatible with antibody stainings.

If anybody tries this with in combination with Drop-seq, I would be interested to hear the outcome. My guess would be it doesn't work - in SCRB-seq, lysates are cleaned up with a Proteinase K digest & RNA desiccation.
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drop-seq, rna integrity, rna isolation, single cell

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