UNIT 2.15 Isolation of Single-Stranded DNA

  1. Yuji Wakimoto,
  2. Jianming Jiang,
  3. Hiroko Wakimoto

Published Online: 1 JUL 2014

DOI: 10.1002/0471142727.mb0215s107

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Wakimoto, Y., Jiang, J. and Wakimoto, H. 2014. Isolation of Single-Stranded DNA. Current Protocols in Molecular Biology. 107:VI:2.15:2.15.1–2.15.9.

Author Information

  1. Department of Genetics, Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 JUL 2014


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. Curr. Protoc. Mol. Biol. 107:2.15.1-2.15.9. © 2014 by John Wiley & Sons, Inc.


  • single-stranded DNA;
  • biotin;
  • streptavidin;
  • magnetic beads