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Old 06-27-2016, 11:08 AM   #6
ATϟGC
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Location: Canada

Join Date: Jun 2013
Posts: 56
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Hello again,

My experience with DNA extraction from RNAlater preserved tissues was mostly plant-related but I am about to do some extractions from RNAlater preserved fish gills so I have been worried about this myself.

A friend was doing some necropsies on 4 Salmo salar the other day and they kindly provided us with some gill tissues so we preserved them in either RNAlater or 100 ETOH for about 24 hours before starting tissue lysis.

I extracted gill tissue from each of the four fish that had been put into the ethanol, RNAlater or RNAlater followed by a one hour PBS bath. This gave a total of 12 gill tissue samples I modified the Xin and Chen method slightly by including proteinase K in the CTAB lysis buffer and made a few other minor modifications.

The tissues immersed in PBS turned into a slimy snot and they were hard to load into the lysis buffer. All the other treatments resulted in a snotty mess at some point of the extraction process (mostly after the chloroform wash but some final eluates had a small snot cloud in the elution buffer).

I ran all the 12 extracts on the nanodrop and the curves were surprisingly nice after seeing so much snot during the extraction. The ETOH samples gave the highest yields and best A260/230 (330 ng/ul, A260/230=2.27) compared to the RNAlater (168 ng/ul, A260/230=1.97) and RNAlater-PBS bathed samples (162 ng/ul, A260/230=1.99). My colleague extracted the same samples with the MN animal tissue kit and got the same general trend.

That is all I intend on doing in the near future with these samples but I'll let you know if my colleague does any PCR or agarose gel with these extracts.

goodluck
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