protocol to use Shrimp DNase - (May/21/2008 )

Hi,

I just received shrimp DNase form USB but they do not send any protocol with it. could any of you give me advices about that ? I'd like to get rid of carry over PCR contamination. My PCR volume is 20µl per reaction. Would 0.1U of enzyme be enough ?

thanks !

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This is the protocol I got for the shrimp nuclease :
You can use it in cDNA synthesis reaction , for that add 1ul into a 20ul cDNA synthesis reaction,running the standard protocol( no activation step requierd) and then to use an inactivation step of 65C for 15min ( or 95C for 10 sec).
If you want to use it after cDNA synthesis step, it`s recommended to use 37C for 15 min and then inactivation step at 65C for 15min.You can use either the buffer from cDNA synthesis step, PCR buffer or make a buffer containning 100mM sodium acetate, pH 5.0 and 5mM MgCl2.
Good luck to you

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thanks !

I'd like to treat the water I use to dilute the DNA templates (of course before mixing up water and DNA). Do you think I need to use the PCR buffer for that ? Or could I just put some shrimp nuclease directly in the water and add MgCl2 ?

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