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Trizol isolation of RNA from OCT-embedded frozen tissue


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#1 cub

cub

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Posted 08 July 2010 - 02:55 PM

Hi all,

I am going to isolate RNA from OCT-Embedded frozen tissue (will cut some slices for this) using Trizol reagent, just wondering if OCT will interfere the process? will OCT affect the yield of RNA? Anyone would provide a detailed protocol will also be greatly appreciated!

Thank you,

Cub

#2 pipetgirl

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Posted 15 December 2010 - 02:28 AM

hello,
did you try it yet? we´re also isolating RNA from cryosections with trizol (in the past we used miRNeasy Kit from QIAGEN and was always successful) but when we are performing more than 20 sections i get a "bubble" on the bottom of the tube after the isopropanol precipitation. the bubble (looks like 20µl of something which doesn´t merge with the isopropanol) moves and there is no pellet visible. when i remove the "bubble" with the isopropanol to continue with the ethanol washing step, my RNA concentrations are very very little.
and the pellet doesn´t dry completely at the end of the protocol and my 260/280 and 260/230 are horrible.

i never had this "bubble" problem in the past and i had no problem to isolate RNA from fresh unfixed tissue or from cellculture. so the trizol should be okay.

any ideas?

ps: i´m using this protocol http://www.scripps.e...on Protocol.pdf but i skip the heating step.
cheers




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