title: Rapid preparation of single stranded DNA from PCR products by streptavidin induced electrophoretic mobility shift.
abstract: 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.
authors.affiliation: NeXstar Pharmaceuticals, Inc., Boulder, CO 80301, USA.
authors.forename: N C
authors.initials: NC
authors.lastname: Pagratis
mesh_terms: D001426:Bacterial Proteins; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D019809:Streptavidin
chemical_list: D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; D019809:Streptavidin