Rapid preparation of single stranded DNA from PCR products by streptavidin induced electrophoretic mobility shift.
AUTOR(ES)
Pagratis, N C
RESUMO
Streptavidin induced electrophoretic mobility shift was used to prepare single stranded (ss) DNA amplified with the polymerase chain reaction in the presence of a biotinylated and a non-biotinylated primer. A variety of denaturing conditions, including incubation at 95 degrees C in 50% formamide can be used without disrupting the streptavidin-biotinylated-ssDNA complex. Following electrophoresis, pure non-biotinylated DNA can be efficiently recovered from 7 M urea gels because it is well separated from the severely retarded streptavidin-biotinylated-ssDNA complex. Quantitative complexing of biotinylated ssDNA can occur at a streptavidin to DNA molar ratio of 1 or more.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=146126Documentos Relacionados
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