denature in SSCP

Mike Morris mike at medsun.unige.ch
Mon Aug 2 12:11:13 EST 1993

In article <1993Aug2.110942.15258 at gserv1.dl.ac.uk>, rmkdhhp at ucl.ac.uk (Dr Huaizheng Peng) writes:
> Hi, group,
> I am doing SSCP at moment. I use routine method to denature DNA fragemnt (95% formamide,0.01%dye, equal volume dilute PCR reaction products,boil 5 min ,and then load on 6-10% gel). I detect dsDNA and ssDNA by Ag staining. I can not denature DNA prope
> PS: I tried add some alkaline into loading mixture, it didnot work.
> Thank you.

I always find that the second time that I do an SSCA ( A for analysis - not 
polymorphism, please!!!) gel, the sample is much better denatured; so, why not
denature - have a coffee - then denature again. Also, try using 2 volumes (not 
one) of formamide/NaOH.


Mike Morris PhD      
Division of Medical Genetics       

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