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Nicking enzyme-based isothermal strand displacement amplification(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2015 06
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Nicking enzyme-based isothermal strand displacement amplification
DENG Shiqiong DONG Juan CHEN Gangyi DU Feng ZOU Jiawei TANG Zhuo
Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China
nicking enzyme isothermal amplification nick strand displacement amplification

Modified substrate (dATPαS) is needed in traditional isothermal strand displacement amplification. In order to realize isothermal strand displacement amplification at conventional substrate, this study replaced the restriction enzyme in traditional isothermal strand displacement amplification with nicking enzyme Nt.BstNBI. The detection limit of plasmid DNA was as low as 2 pmol/L. The amplification reaction could be completed in 15-30 min, with products directly detectable by agarose gel electrophoresis. The results showed that the nicking enzyme-based isothermal strand displacement amplification is an environment-friendly, cost effective new technology that is faster, more convenient, and practical than the traditional method, especially for on-site testing.


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Last Update: 2016-01-05