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Difficulty in cutting tissue after collection of sample in RNAlater


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#1 Afiqah Alyaa

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Posted 14 September 2012 - 01:29 AM

I kept my tissue samples in RNAlater after collection from the OT.
The samples were kept overnight in 4 degrees, and transferred to -80 degrees the next day.

When trying to cut the tissue samples with cryostat,
the samples seem to split and break and I cannot prepare my slides with this samples.
Before subjecting my samples to the OCT medium, I just dab the samples on a tissue paper to get rid of the RNAlater.

So do the difficulty in cutting the tissue samples are caused by RNAlater ?
How to remove the RNAlater covering the tissue sample effectively before subjecting them for slide preparation ?

Hope that someone out there can help me to overcome this problem.

Thank you.

#2 Curtis

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Posted 14 September 2012 - 08:13 PM

RNAlater is meant to save RNA from degradation. It is not mean to keep the tissue at its original shape. We always cut our sample, save some of it in RNAlater, and keep the rest in other buffers.

#3 bob1

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Posted 14 September 2012 - 09:46 PM

Yeah, RNAlater is mostly a sulphate, which is used to precipitate proteins, not preserve them. If you want to preserve tissue properly, use a fixative such as formaldehyde.

#4 Afiqah Alyaa

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Posted 16 September 2012 - 06:12 AM

Thanks guys for your comments.

I have just came across an article that mentioned to incubate the tissue collected in RNAlater in ice-cold PBS for 5-10mins before cutting them using cryostat.




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