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FW: Ethanol in my RNA/TE buffer

FW: Ethanol in my RNA/TE buffer - Protocols and Methods Forum

FW: Ethanol in my RNA/TE buffer - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 06-23-2004, 08:03 PM
Deanne Bell
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Default FW: Ethanol in my RNA/TE buffer



Adding ethanol (like 250% of the buffer volume) makes nucleic acids
precipitate out of buffered solution - right?
A LOT of left over ethanol will prevent nucleic acids from fully
dissolving back into the buffer. A LITTLE will probably evaporate out
of the TE solution.

TTFN
Dee Bell



-----Original Message-----
From: [Only registered users see links. ] [mailtowner-methods@hgmp.mrc.ac.uk]
On Behalf Of Mark Ledesma
Subject: Ethanol in my RNA/TE buffer
I know it is customary when washing an RNA pellet with 75% ethanol to
dry the ethanol off the pellet before suspending it in TE buffer.
Could someone kindly explain why?

Someone I know who looks kind of like me (it isn't me, really) just
forgot to dry his pellet and surely has some ethanol in with his RNA
and TE. How bad is this?

Thanks for any help,
Mark Ledesma
---
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  #2  
Old 06-25-2004, 12:07 AM
redeamer
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Default FW: Ethanol in my RNA/TE buffer

Hi Deanne!


I prefer to precipitate the RNA with 2.5 volumes of isopropanol : 5M
NH4OAc (5:1) at room temperature. That is much more effective than
ethanol one. Afterwards, i wash the transparent RNA pellet with 70%
EtOH.


Well, if you saw the pellet and didn't use any coprecipitant then you
have quite a lot of RNA. Residual EtOH won't heart you if the
applications (assays) do not require much RNA, say if you dilute RNA
20 fold or 50 fold. Nothing to worry about. Typically it is not a
problem.

But if you think or have intrinsic feeling that you should actually
dry it completely. Just reprecipitate the RNA



---> Drew
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  #3  
Old 06-25-2004, 12:07 AM
redeamer
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Posts: n/a
Default FW: Ethanol in my RNA/TE buffer

Hi Deanne!


I prefer to precipitate the RNA with 2.5 volumes of isopropanol : 5M
NH4OAc (5:1) at room temperature. That is much more effective than
ethanol one. Afterwards, i wash the transparent RNA pellet with 70%
EtOH.


Well, if you saw the pellet and didn't use any coprecipitant then you
have quite a lot of RNA. Residual EtOH won't heart you if the
applications (assays) do not require much RNA, say if you dilute RNA
20 fold or 50 fold. Nothing to worry about. Typically it is not a
problem.

But if you think or have intrinsic feeling that you should actually
dry it completely. Just reprecipitate the RNA



---> Drew
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buffer , ethanol , rna or te


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