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TRIzol RNA extraction- What is in the top layer?


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#1 egonzalez

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Posted 21 February 2011 - 03:21 AM

Hello,

I am doing Arabidopsis RNA extraction with the TRIzol reagent, adding RNA plant isolation aid (http://www.ambion.co...CatNum.php?9690).

After I add TRIzol reagent to the grinded tissue (5' in ice) then I add this plant isolation aid and centrifuge for 5'.
In the centrifuged tubes I have the pellet with tissue debris at the bottom, the aquous phase with RNA and on top I have another layer of a different colour, a bit blurry, and I wanted to know what is made of and more importantly if I should take it when I get the supernatant to the next step in the protocol.

Thanks in advance!

#2 Mary K

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Posted 25 February 2011 - 01:38 PM

Hello,

I am doing Arabidopsis RNA extraction with the TRIzol reagent, adding RNA plant isolation aid (http://www.ambion.co...CatNum.php?9690).

After I add TRIzol reagent to the grinded tissue (5' in ice) then I add this plant isolation aid and centrifuge for 5'.
In the centrifuged tubes I have the pellet with tissue debris at the bottom, the aquous phase with RNA and on top I have another layer of a different colour, a bit blurry, and I wanted to know what is made of and more importantly if I should take it when I get the supernatant to the next step in the protocol.

Thanks in advance!


I think this is lipds or maybe waxes from the cutical layer of the plant.I would not take it. I see this same layer when I extract RNA from insect larvae. In the case of the larvae, I know it's lipids because when I first saw it I touched it - it's oily!! I would leave it behind.




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