Hi everyone,
I am currently working on RNA extraction on tissue, but unfortunately, I only have large frozen clinical tissue on hand.
In order to maximize the RNA integrity, I plan to use cryostat to shave the amount of tissue I need. However, I am afraid that OCT mounting compound may affect the downstream RNA extraction and I also want to keep the remaining tissue sample still usable for other omic analysis.
So, can DEPC-treated water be used as a replacement for the OCT compound as a mounting media? if so, how can I mount the large tissue (~2cm*2cm*1cm) on the mounting disk with DEPC-treated water in the cryostat?
Also, will be very much appreciated if there is another method that can prevent tissue from thawing during cutting other than using crostat.
Thanks!
The OCT can be a problem in RNA extraction. I suggest embedding in sucrose to prevent ice crystal lysing of cells and shattering tissue instead of recoverably shaving it: https://cells.ucsc.edu/cortex-dev/protocol.pdf
Alternatively, the amount of OCT can be minimized: https://kb.10xgenomics.com/hc/en-us/articles/360053675712-Can-OCT-affect-bulk-RNA-extraction-and-RIN-assessment
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