DEPC treatment of ethanol
Ali El-Salanti
precipitation. If yes how do you autoclave 100% ethanol.
Thanks
AS
Frank Ludewig
mix DEPC-treated water and ethanol (p.a.) to yield 70 % ethanol.
Bye, FrankL
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University of Cologne
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50931 Cologne
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email: frank....@uni-koeln.de
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Ali El-Salanti <isl4...@worldonline.dk> schrieb in im Newsbeitrag:
8uo79e$r90$1...@news.net.uni-c.dk...
marianne carter
Same as you dont DEPC treat other solutions you make up eg EDTA
etc........you use DEPC treated water only as it breaks specific chemical
bonds ie heat treatment of some chemical solutions for use inn RNA
purification.
"Ali El-Salanti" <isl4...@worldonline.dk> wrote in message
news:8uo79e$r90$1...@news.net.uni-c.dk...
Ali El-Salanti
water? But what if the things I use are not "ultra pure - high grade RNAse
free" fx EDTA or sodium acetate?
Thanks
Ali
"marianne carter" <mcar...@optusnet.com.au> wrote in message
news:3a0fc68b$0$19422$7f31...@news01.syd.optusnet.com.au...
Rogier
> So when I make buffer I do not have to DEPC treat it as long as I use
DEPC
> water?
Yeah, you gotta DEPC everything. That's why most RNA people use HEPES
instead of Tris (HEPES can be DEPCed, Tris not). DEPC reacts with
amines, so anything that contains amines (Tris, EDTA) can not be DEPCed.
No need to DEPC ethanol, since RNases are not soluble in pure alcohol.
If you're really paranoid, you can spin big tubes of ethanol to make
sure any RNase in there is removed.
(as you know, even denatured RNases can be revived when mixed with
water. you can extract active RNase A from the white interphase you get
in phenol extractions...)
Good luck,
Rogier
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Rogier Stuger
Dept MicFizz, Free U of A
E rogier AT biogate DOT com (spammers will be killed)
pleaz do NOT use the my-deja.com address. I never read that one
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Before you buy.
Michael L. Sullivan
>Same as you dont DEPC treat other solutions you make up eg EDTA
>etc........you use DEPC treated water only as it breaks specific chemical
>bonds ie heat treatment of some chemical solutions for use inn RNA
>purification.
>
DEPC CAN be used to treat some solutions, just not those made with
amine-containing compounds, like TRIS. However, LiCl, NaOAc solutions and
the like can be DEPC treated, although such precautions are often not
needed if care is taken, even for water. As for the specific question
regarding treatment of EtOH-- just use an unopened bottle and use
DEPC-treated (or other RNAse free source) water to make the 70%.
Mike
Michael L. Sullivan, Ph.D
U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706
(608) 264-5144 Phone
(608) 264-5147 Fax
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Dr. Hiranya S. Roychowdhury
Absolutely don't try to "treat" Etoh w/ DEPC! How do you propose to get rid
of DEPC after you have added it to the etoh?
At 09:03 AM 11/13/00 +0100, Ali El-Salanti wrote:
>Is it necessary to DEPC treat 70 and 100% ethanol prior to RNA
>precipitation. If yes how do you autoclave 100% ethanol.
>Thanks
>AS
>
>
>
>
Dr. Hiranya Sankar Roychowdhury
College Asst. Prof.
Molecular Biology,
Dept. of Chemistry & Biochemistry
Box 30001 - 3MLS
New Mexico State University
Las Cruces, NM 88003
Lab: (505) 646 4722
Office: (505) 646 8256
hroy...@nmsu.edu
---
Ali El-Salanti
""Michael L. Sullivan"" <mlsu...@facstaff.wisc.edu> wrote in message
news:v04011700b635acd47b24@[144.92.64.179]...
Michael L. Sullivan
>
Well, the DEPC simply reacts with the excess of amine groups in solution.
Consequently, the DEPC is not available to react with amine groups in
proteins, like RNase. So in this case, it's not a matter of something bad
happening, just a case of wasting money on a treatment that would be
ineffective.
Gauthier , Eric
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