RNase Zap (Invitrogen) and RNase Away (Molecular Bioproducts)
Igor Sagdeev
RNase Zap (Invitrogen)
and
RNase Away (Molecular Bioproducts)?
Thank you in advance!
Christian Praetorius
>The "RNAses are so
>everywhere that the only way to get rid of them is to treat everything
>with some magic expensive solutions" is basically a myth.
Can you tell this our technician?
Christian
--
X-no-Sig: yes
talulah gosh
Michael Sullivan
On Feb 16, 2009, at 6:16 PM, DK wrote:
> In article <mailman.225.123480...@net.bio.net>, Igor
> Sagdeev <sag...@gmail.com> wrote:
>> Can anyone comment of the difference(s) between these products:
>>
>> RNase Zap (Invitrogen)
>>
>> and
>>
>> RNase Away (Molecular Bioproducts)?
>
> Well,
>
> MSDS for both products makes it reasonably obvious that RNAse Away
> is simply a sodium hydroxide at unspecified concentration and RNase
> Zap is simply an SDS at unknown concentration.
>
> Take your pick. I'd contend that with already clean glassware that was
> not handled by bare hands neither is necessary. The "RNAses are so
> everywhere that the only way to get rid of them is to treat everything
> with some magic expensive solutions" is basically a myth.
>
> DK
> _______________________________________________
>
The more one works with RNA, the one more realizes that DK's point is
true!
My favorite example of the extreme paranoia of RNases was a lab I
knew of that baked the mortars and pestles they used to grind tissue
for RNA preps! So there might be a little RNase on those: far more
will be liberated when the tissue gets ground! They did go through a
lot of mortars since these tended to break when cooling too fast out
of the oven.
As for magic solutions, I personally use dilute bleach when I feel
some treatment is called for. Ambion's web site even says this is an
alternative to their solution.
Another hint is that usually there is no special need to treat water
with DEPC. My experience is that 18 megaohm water from a milliQ
system is essentially RNase free.
Mike
---
Michael L. Sullivan
Plant Research Molecular Geneticist
US Dairy Forage Research Center
ARS-USDA
1925 Linden Drive West
Madison, WI 53706
(608) 890-0046 (Phone)
(608) 890-0076 (FAX)
Dr. Hiranya S. Roychowdhury
baking the glasswares always worked rather well. Of course, whoever pours
LiqN2 on the Mortar/pestle straight out of the oven has a problem far more
serious than RNase in the prep. ;)
Hiranya.
>
> On Feb 16, 2009, at 6:16 PM, DK wrote:
>
>> In article <mailman.225.123480...@net.bio.net>, Igor
>> Sagdeev <sag...@gmail.com> wrote:
>>> Can anyone comment of the difference(s) between these products:
>>>
>>> RNase Zap (Invitrogen)
>>>
>>> and
>>>
>>> RNase Away (Molecular Bioproducts)?
>>
> _______________________________________________
> Methods mailing list
> Met...@net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>
--
Hiranya S. Roychowdhury, Ph.D.
Asst. Professor,
Health & Public Services
Dona Ana Community College
New Mexico State University
Las Cruces, NM 88003
Aawara Chowdhury
DK <d...@no.email.thankstospam.net> wrote:
> Take your pick. I'd contend that with already clean glassware that was
> not handled by bare hands neither is necessary. The "RNAses are so
> everywhere that the only way to get rid of them is to treat everything
> with some magic expensive solutions" is basically a myth.
Let me elaborate on this statement, because I take issue with it in part.
My Ph.D. thesis was on the structure of retroviral RNAs. The structural
analysis, obtained in part by digestion with structure/sequence specific
RNases of end-labeled viral RNAs, was exquisitely sensitive to nuclease
contamination. I've recovered RNase A activity in bottles that were
autoclaved .....
Having said that, for 99% of uses, the background level of RNase makes
no difference at all. Further, while I can appreciate that there may
be a loss of signal if one does a Northern for full-length mRNA, these
days, most quantitation is performed using techniques such as real-time
PCR of reverse-transcribed samples. Such techniques are much less
sensitive to small amounts of degradation by RNases in the environment.
AC
--
Email: echo 36434455860060025978157675027927670979097959886449930P | dc
Christian Praetorius
>The more one works with RNA, the one more realizes that DK's point is
>true!
Its important to work careful and clean. But this applies to all lab
work, which is done carefully.
>Another hint is that usually there is no special need to treat water
>with DEPC. My experience is that 18 megaohm water from a milliQ
>system is essentially RNase free.
Try to bring this into the minds of people.
Christian
--
X-no-Sig: yes
Jayakumar, R
But RNAse contamination of the milliq water can happen when it is in
transit to your reaction tubes which may also be RNAse free, and also
the tubing which dispenses the water from the Milliq cartridge to your
"Rnase-free" storage container. It is important the glass bottles or
carboys or other plasticware that is used to store this milliQ water is
clean too, wherein lies the problem that it is virtually impossible to
eliminate RNAses during the transitionary phase. That is why it is
always PRUDENT (an important word in the vocabulary of good scientists)
to treat the water with DEPC in the bottle it is going to be stored,
hence eliminating all external sources of RNAse. Anyone care to
comment?? And I don't see any reason why people should try to save a
few minutes of their time and a few dollars and risk their expensive
experiments.
Jay
-----Original Message-----
From: methods...@oat.bio.indiana.edu
[mailto:methods...@oat.bio.indiana.edu] On Behalf Of DK
Sent: Wednesday, February 18, 2009 12:52 AM
To: met...@magpie.bio.indiana.edu
Subject: Re: RNase Zap (Invitrogen) and RNase Away (Molecular
Bioproducts)
In article <mailman.233.123490...@net.bio.net>, Michael
Sullivan <mlsu...@wisc.edu> wrote:
>
>Another hint is that usually there is no special need to treat water
>with DEPC. My experience is that 18 megaohm water from a milliQ system
>is essentially RNase free.
Of course. The water goes through several cartridges that all absorb
proteins and peptides (ion exchange resins, activated charcoal and even
the final 0.2 micron filter that is not designed to be low protein
binding) plus the whole system is "sanitized" with NaOH on a periodic
basis, killing any microbes that might be a source of RNAses.
DK
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Nick Theodorakis
wrote:
> But RNAse contamination of the milliq water can happen when it is in
> transit to your reaction tubes which may also be RNAse free, and also
> the tubing which dispenses the water from the Milliq cartridge to your
> "Rnase-free" storage container. It is important the glass bottles or
> carboys or other plasticware that is used to store this milliQ water is
> clean too, wherein lies the problem that it is virtually impossible to
> eliminate RNAses during the transitionary phase. That is why it is
> always PRUDENT (an important word in the vocabulary of good scientists)
> to treat the water with DEPC in the bottle it is going to be stored,
> hence eliminating all external sources of RNAse. Anyone care to
> comment?? And I don't see any reason why people should try to save a
> few minutes of their time and a few dollars and risk their expensive
> experiments.
My comment is the same as the others who note that ultra-pure water is
free of RNase contamination and that RNAse-phobia is way overdone. I
haven't used any DEPC or similar reagents since grad school, and
haven't had a problem even when probing for full length mRNAs in
Northerns. I never understood why anyone would pay good money to get
their water ultrapure and then deliberately contiminate with something
else. If your water is not pure enough to use for RNA, then it's
probably not good for most other applications.
I don't store water for RNA use, in any case; I get it fresh from the
tap.
Nick
--
Nick Theodorakis
nick_the...@hotmail.com
contact form:
http://theodorakis.net/contact.html
Michael Sullivan
an experiment over which you have little control than the brief time
you will have a container open to dispense water from the milliQ into
it. How long will a bottle or tube of DEPC treated water be open
while you are later dispensing it into reaction tubes, how long are
reaction tubes open while you are adding other reagents, etc? And how
about all those other reagents that go into an experiment, many of
which you cannot treat with DEPC? And DEPC treating the water because
the container might not be clean enough seems extreme since glassware
could be treated with dilute base or bleach or baked to render it
RNase free, or one could use sterile disposable plasticware which is
again, in my experience, RNase free whether marketed that way or not.
Also, where do you draw the line at being "prudent"? Besides gloves,
should you wear a surgical cap, gown and mask? Should you work in a
hood or a glove box?
In my experience, in a reasonably clean lab, there just isn't RNase
all over the place ready to fall into your experiment and using
MilliQ water directly without DEPC treatment has simply not been a
problem for me or (judging from previous discussions here) many other
researchers. So while I agree we shouldn't be penny wise and pound
foolish, I don't have a problem saving my pennies by not spending
them to solve a non-existent problem.
Mike
On Feb 18, 2009, at 10:55 AM, Jayakumar, R wrote:
>
> But RNAse contamination of the milliq water can happen when it is in
> transit to your reaction tubes which may also be RNAse free, and also
> the tubing which dispenses the water from the Milliq cartridge to your
> "Rnase-free" storage container. It is important the glass bottles or
> carboys or other plasticware that is used to store this milliQ
> water is
> clean too, wherein lies the problem that it is virtually impossible to
> eliminate RNAses during the transitionary phase. That is why it is
> always PRUDENT (an important word in the vocabulary of good
> scientists)
> to treat the water with DEPC in the bottle it is going to be stored,
> hence eliminating all external sources of RNAse. Anyone care to
> comment?? And I don't see any reason why people should try to save a
> few minutes of their time and a few dollars and risk their expensive
> experiments.
>
---
Christian Praetorius
>In my experience, with autoclaves being dirty, dirty things, anything
>autoclaved is simply not clean. Sterile - yes, but not clean. Few
Especially, when the same autoclave is used for autoclaving trash and
clean things.
>lab it is a rule that protein crystallization has to be done with tips
>out of the bag, never to be autoclaved.
These tips (of course not touched with bare fingers) should be free of
everything because of the production process.
Christian
--
X-no-Sig: yes
Aawara Chowdhury
DK <d...@no.email.thankstospam.net> wrote:
> Aawara, I don't doubt your observation for a nanosecond but let
> me ask this: Any chance that the RNAse activity was there
> *because* of the autoclaving?
100% possible.