Optimizing PCR Reactions

Ali Karami ali at biobase.aau.dk
Mon Jan 24 09:06:51 EST 1994


In <2hbvpk$rpg at eldborg.rhi.hi.is> php at rhi.hi.is (Petur Henry Petersen) writes:

  >In <2h4kkn$kbl at mserv1.dl.ac.uk> "" <unknown at dl.ac.uk> writes:

>>Hi-
>>I'm trying to find little tricks for optimizing PCR reactions.  I read a
>>note in Trends In Genetics which mentioned 5-10% DMSO would help by
>>reducing secondary structure, but the article also said that DMSO could
>>inhibit Taq.  Does anyone have any experience with this?  It didn't work
>>for me. 

>>Aside from playing with primer/template ratio and salt concentration, what
>>else can be done to optimize a PCR reaction?   At least one biotech company
>>sells an optimization kit - any ideas what's in it?

>>Thanks,
>>Robert Schoenfeld
>>Univ. Utah
>>schoenfeld at msscc.med.utah.edu

>	* heat
>	* time
>	* try using dummy vials in pcr (TIG May 1991 vol 7 no. 5)
>	* Try using Rnase on the DNA
>	* concentration of Taq 
>	* use some other kind of Taq
>	* other buffers
>	* hotstart methods
>	* concentration of dNTP
>	* Concentration of Mg
        * Concentration of Primers

>	Please add to this list if you can!


>	P.Henry
>	U.Iceland
>	php at lif.hi.is




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