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How to make nuclease free water?


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#1 pygo

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Posted 29 March 2005 - 08:31 PM

Hi ya all!

I'm looking for a way to make my own nuclease free water for RT-PCR. I know you can buy it nowadays, but in my country it's gonna be a bit difficult. Can anyone suggest what water treatment I should have or is autoclaving the MilliQ water is good enough?

Thanx a lot

Sissle

#2 ajames

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Posted 29 March 2005 - 09:48 PM

Autoclaving will not remove RNase contamination. The method to use is to add DEPC to the water and then autoclave. The autoclaving degrades the DEPC which would interfere with later reactions. I understand that it may be difficult to get PCR grade water but is probably worth the effort since DEPC is a neurotxin and is often still present even after autoclaving. Is PCR grade water really any more difficult to source than any of the other PCR reagents? Remember that the PCR will only be as good as your worst reagent.

#3 fred_33

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Posted 30 March 2005 - 02:33 AM

hi
this topic may help you too :
http://www.protocol-...?showtopic=5810




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