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Old 07-19-2018, 05:06 AM   #1
thiNGS
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Location: Netherlands

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Default stabilize the RNA expression profile upon harvesting samples

Hi all. I am designing an experiment for RNAseq. I start treating my cells with a compound (or DMSO in the control) and collect samples at different timepoints. My question is: when I harvest the cells centrifuging for 10 minutes I am pretty sure the expression is going to slightly change and I would like to avoid that. I have read about using RNAprotect adding it directly to the culture but I have a feeling this reagent will just inhibit RNases to increase RNA stabilisation but it won't really "freeze" the expression profile at that particular timepoint. Any suggestions?
Thank you in advance
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Old 07-19-2018, 11:42 AM   #2
kc3228
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Your best bet is probably to add trizol or similar directly to the plates and scrape the cells off, then extract RNA. Other than that, try to keep things cold.
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