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Helicase‐dependent isothermal DNA amplification
Author(s) -
Vincent Myriam,
Xu Yan,
Kong Huimin
Publication year - 2004
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/sj.embor.7400200
Subject(s) - dna polymerase , multiple displacement amplification , primer (cosmetics) , dna , polymerase , loop mediated isothermal amplification , dna clamp , microbiology and biotechnology , applications of pcr , helicase , primase , polymerase chain reaction , biology , chemistry , genetics , multiplex polymerase chain reaction , gene , rna , reverse transcriptase , dna extraction , organic chemistry
Polymerase chain reaction is the most widely used method for in vitro DNA amplification. However, it requires thermocycling to separate two DNA strands. In vivo , DNA is replicated by DNA polymerases with various accessory proteins, including a DNA helicase that acts to separate duplex DNA. We have devised a new in vitro isothermal DNA amplification method by mimicking this in vivo mechanism. Helicase‐dependent amplification (HDA) utilizes a DNA helicase to generate single‐stranded templates for primer hybridization and subsequent primer extension by a DNA polymerase. HDA does not require thermocycling. In addition, it offers several advantages over other isothermal DNA amplification methods by having a simple reaction scheme and being a true isothermal reaction that can be performed at one temperature for the entire process. These properties offer a great potential for the development of simple portable DNA diagnostic devices to be used in the field and at the point‐of‐care.