Unit

UNIT 15.11 Helicase-Dependent Amplification of Nucleic Acids

  1. Yun Cao,
  2. Hyun-Jin Kim,
  3. Ying Li,
  4. Huimin Kong,
  5. Bertrand Lemieux

Published Online: 11 OCT 2013

DOI: 10.1002/0471142727.mb1511s104

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Cao, Y., Kim, H.-J., Li, Y., Kong, H. and Lemieux, B. 2013. Helicase-Dependent Amplification of Nucleic Acids. Current Protocols in Molecular Biology. 104:15.11:15.11.1–15.11.12.

Author Information

  1. BioHelix Corporation, Beverly, Massachusetts

Publication History

  1. Published Online: 11 OCT 2013

Abstract

Helicase-dependent amplification (HDA) is a novel method for the isothermal in vitro amplification of nucleic acids. The HDA reaction selectively amplifies a target sequence by extension of two oligonucleotide primers. Unlike the polymerase chain reaction (PCR), HDA uses a helicase enzyme to separate the deoxyribonucleic acid (DNA) strands, rather than heat denaturation. This allows DNA amplification without the need for thermal cycling. The helicase used in HDA is a helicase super family II protein obtained from a thermophilic organism, Thermoanaerobacter tengcongensis (TteUvrD). This thermostable helicase is capable of unwinding blunt-end nucleic acid substrates at elevated temperatures (60° to 65°C). The HDA reaction can also be coupled with reverse transcription for ribonucleic acid (RNA) amplification. The products of this reaction can be detected during the reaction using fluorescent probes when incubations are conducted in a fluorimeter. Alternatively, products can be detected after amplification using a disposable amplicon containment device that contains an embedded lateral flow strip. Curr. Protoc. Mol. Biol. 104:15.11.1-15.11.12. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • isothermal amplification;
  • nucleic acid quantitation;
  • lateral flow strip