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.

Keywords: biotin; magnetic beads; single-stranded DNA; streptavidin.