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Plant RNA Isolation Aid


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#1 kaluts

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Posted 20 December 2009 - 06:59 PM

I'm using a Plant RNA Isolation Aid from Ambion to extract RNA from wood samples. The problem is my supernatant turns cloudy, which clogs the filters. Any ideas how to work around this and why does this is happen in the first place? Any advice is GREaTLY appreciated.

#2 gogreen

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Posted 21 December 2009 - 09:57 AM

What kit are u using? the Ambion isolation kits combined with Ambion RNA isolation aid?? pls post more details

#3 molgen

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Posted 21 December 2009 - 11:43 PM

In my experience, the plant RNA isolation kits only work on things like tomato leaf or Arabidopsis.
For woody parts the kits don't work manly because thy are rich with lignin and suberin.
The cloudiness you mention probably is one of them becoming unsalable and that's why they clog your filters.
I'm sad to say that most probably you will have to use a CTAB protocol preferably with PVP.

#4 kaluts

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Posted 23 December 2009 - 08:08 PM

In my experience, the plant RNA isolation kits only work on things like tomato leaf or Arabidopsis.
For woody parts the kits don't work manly because thy are rich with lignin and suberin.
The cloudiness you mention probably is one of them becoming unsalable and that's why they clog your filters.
I'm sad to say that most probably you will have to use a CTAB protocol preferably with PVP.


Guys, i got somethings mixed up. Sorry about that. I used Plant RNA Isolation Aid(Ambion) to extract RNA from my wood samples. These samples turned out fine. However, i started sampling from older trees of the same species. The supernatant from the new samples were clear when i used the Plant Kit. Next, I used a RNAqueous kit to purify my RNA. My supernatant turned cloudy when i put in 64% ethanol. When this goes through the filter, it clogs. Does anyone has any ideas on this?




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