About SSCP

[Glenn Dunshea]

I have found MDE solution excellent for detection of mutations without the
need for glycerol. Something to consider when running your trials is that
one of the most critical parameters for SSCP assays is constant temperature.
I would not recommend running SSCP at room temperature..
I hope your experiments are successful.
Cheers,
Glenn

On Thu, Jul 30, 2009 at 7:49 AM, Marcos Moraes <markydor from yahoo.com.br>wrote:

> Dear Bruno,
>
> Surfing on net I found a message (
> http://www.bio.net/bionet/mm/methods/2002-February/092258.html) that
> you said things about SSCP resolution. In that moment, you said you was
> running 1XTBE + 10% glycerol gels. How many conditions have you ever tested?
> I'm asking because after a long time screening for mutation for COL1A1 gene
> (Osteogenesis imperfecta), i realized that the low ratio of mutants could be
> the SSCP conditions. Here, the SSCP runs at room temperature (PAGE-SSCP S2
> Biometra) and at the moment, I'm trying to figure out a condition(s) capable
> to detect the mutations. I'm testing absence, 5% and 10% glycerol; no
> glycerol + 10% sucrose; MDE 0.5X w/ or w/out glycerol 5%, all of them with
> 0.5X TBE. Do you would recommend me to use 1X TBE instead of 0.5X? Does it
> make any difference for detection?
>
> Best regards,
>
> Marcos Moraes| Mestrando em Biotecnologia | Saúde
> Laboratório de Genética Humana e Molecular
>
> Núcleo de Genética Humana e Molecular | Depto. Ciências Biológicas | Centro
> de Ciências Humanas e Naturais | Universidade Federal do Espírito Santo
> Av. Marechal Campos, 1468 - Maruípe - 29040-090 - Vitória - ES - Brasil
> T +55 27 3335 7253 | C+55 27 9249 0190 | markydor from yahoo.com.br
> http://lattes.cnpq.br/0799432282422121
>
>
>
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