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J Biochem. 2007 Aug;142(2):273-81. Epub 2007 Aug 24.

Highly efficient isothermal DNA amplification system using three elements of 5'-DNA-RNA-3' chimeric primers, RNaseH and strand-displacing DNA polymerase.

Author information

1
Products Development Center, Takara Bio Inc., 2257, Noji, Kusatsu, Shiga 525-0055, Japan.

Abstract

We developed an efficient method of isothermally amplifying DNA termed ICAN, Isothermal and Chimeric primer-initiated Amplification of Nucleic acids. This method allows the amplification of target DNA under isothermal conditions at around 55 degrees C using only a pair of 5'-DNA-RNA-3' chimeric primers, a thermostable RNaseH and a DNA polymerase with strong strand-displacing activity. ICAN is capable of amplifying DNA at least several times greater than the amount produced with PCR by increasing primer concentration. This method would be applicable for on-site DNA detection including gene diagnosis, and would also be suitable for 'real time' detection when combined with a cycling probe.

PMID:
17720718
DOI:
10.1093/jb/mvm138
[Indexed for MEDLINE]

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