TRIzol RNA extraction- What is in the top layer? - (Feb/21/2011 )
Hello,
I am doing Arabidopsis RNA extraction with the TRIzol reagent, adding RNA plant isolation aid (http://www.ambion.com/catalog/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!
-egonzalez-
egonzalez on Mon Feb 21 11:21:20 2011 said:
Hello,
I am doing Arabidopsis RNA extraction with the TRIzol reagent, adding RNA plant isolation aid (http://www.ambion.com/catalog/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.
-Mary K-