RNase Zap (Invitrogen) and RNase Away (Molecular Bioproducts)

Michael Sullivan via methods%40net.bio.net (by mlsulliv from wisc.edu)
Wed Feb 18 16:03:11 EST 2009


It seems to me there is far more "transitory" time in the course of  
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.
>
> Jay
>
> -----Original Message-----
> From: methods-bounces from oat.bio.indiana.edu
> [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of DK
> Sent: Wednesday, February 18, 2009 12:52 AM
> To: methods from magpie.bio.indiana.edu
> Subject: Re: RNase Zap (Invitrogen) and RNase Away (Molecular
> Bioproducts)
>
> In article <mailman.233.1234903817.13724.methods from net.bio.net>, Michael
> Sullivan <mlsulliv from 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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---
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)



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