title: Isolation of single-stranded DNA.
abstract: Single-stranded DNA (ssDNA) is often used for DNA sequencing as well as microarray and hybridization technologies. Asymmetric PCR or exonuclease digestion followed by urea gel separation and isolation on streptavidin-coated magnetic beads are commonly used for this purpose. These two methods may not yield large amounts of highly purified ssDNA. This protocol for ssDNA isolation from PCR-amplified DNA involves biotin labeling of one strand and subsequent strand separation utilizing streptavidin-coated magnetic beads.
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Wakimoto
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Jianming
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Wakimoto
mesh_terms: D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000302:isolation & purification*; D060526:Magnetic Fields; D016133:Polymerase Chain Reaction / Q000379:methods; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D004277:DNA, Single-Stranded; D019809:Streptavidin
keywords: biotin; magnetic beads; single-stranded DNA; streptavidin