About SSCP

Marcos Moraes via methods%40net.bio.net (by markydor from yahoo.com.br)
Wed Jul 29 18:49:14 EST 2009


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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