Does anybody have experience with RNA extraction from animal tissues preserved in RNAlater (Qiagen and Ambion)? We want to extract RNA from animal tissues stored in RNA later. Suppose if we use Qiagen RNeasy tissue kit or TRizol method for RNA extraction, do we need to remove the RNAlater by washing tissues in PBS or some other stabilizing solution? Does anybody have experience in this method?
I washed tissue stored RNAlater in PBS, then proceeded with RNA extraction. RIN increased from 2 (without washing) to 8-9.
You can find the details how to remove the RNA later from animal tissues in the Ambion pdf file at pag 8:
http://www.invitrogen.com/etc/medialib/en/filelibrary/Nucleic-Acid-Purification-Analysis/PDFs.Par.18819.File.dat/bp_7020.pdf
Please refer to the article published by Mutter et al 2004, BMC Genomics "Comparison of frozen and RNALater solid tissue storage methods for use in RNA expression microarrays" (Methods section). Hope it helps......
The tissues are dried by pressing them between clean wipes to remove any RNA later before any further step of extraction.
I washed tissue stored RNAlater in PBS, then proceeded with RNA extraction. RIN increased from 2 (without washing) to 8-9.
I agree with Agnes, but please remove RNAlater as much as you can because any traces will interfere with the subsequent processing of Trizol extraction.
So if am using RNAlater and later RNeasy mini kit, both from qiagen do i still need to remove RNAlater?
In the QIAGEN manual for storage of tissue at -80 it says to first incubate at 2-8 degrees and then remove the tissue form the reagent for storage at -80.
Why would the removal of the tissue from the solution be important in this procedure??? why not store it at -80 in the RNA later solution?
storage is not a problem. i was thinking of RNA extraction. wouldn't it interfere when we are trying to extract RNA from this tissue?
I have the same question, i have tissues in rnalter for around 2 years. Before extraction, i sequeeze them to remove rnalater. Is it necessary to do this? Or we can proceed as I am using qiagen rneasy mini kit. Any thoughts ?
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