concentrated dNTPs

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Mon Nov 30 04:31:44 EST 1998


In article <73r69t$c1i$1 at news.worldonline.nl>, J. Hendriks
<hendrikb at worldonline.nl> writes
>
>vinsin at my-dejanews.com wrote: <73r164$n5d$1 at nnrp1.dejanews.com>...
>>Dear friends,
>>
>>I have to do some PCR reactions in which I have to use dNTPs of the
>>concentraion 200mM. I am not getting address of the supplier from where I
>can
>>procure the same. Kindly help me out from the fix.
>>
>>Thanks in advance.
>>
>>VINAY
>>
>>-----------== Posted via Deja News, The Discussion Network ==----------
>>http://www.dejanews.com/       Search, Read, Discuss, or Start Your Own
>
>Are you sure about this concentration? It seems a awfull lot to me (I use in
>my pcr-reactions an endconcentration dNTP's of 0.2 micromolar). By using
>this amount of dNTP's the Taq-polymerase will have a very hard time doing
>anything at all.

If you try to use 200mM dNTPs (final concn.) in a PCR it will not work
due to them chelating out all the Mg in the reaction. Try 0.2mM final ie
2ul of a 5mM dNTP mix (all 4 together) in a 50ul PCR.

If you are trying to make 200mM stocks then you will have to buy solid
and reconstitute and don't forget to pH to 7.0. The highest I've ever
seen commercial dNTPs sold at is 100mM although they will dissolve to
250mM.

Duncan 
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk



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