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DNA Detection by Cascade Enzymatic Signal Amplification

  • Bingjie Zou
  • Yinjiao Ma
  • Guohua Zhou
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1039)

Abstract

Although many approaches based on template replication were developed and applied in DNA detection, cross-contamination from amplicons is always a vexing problem. Thus, signal amplification is preferable for DNA detection due to its low risk of cross-contamination from amplicons. Here, we proposed a cascade enzymatic signal amplification (termed as CESA) by coupling Afu flap endonuclease with nicking endonuclease, including three steps: invasive signal amplification, flap ligation, and nicking endonuclease signal amplification. Because of the advantages of low risk of contamination, no sequence requirement of target DNA, and the universal reaction conditions for any target detection, CESA has a great potential in clinical diagnosis.

Key words

Signal amplification Afu flap endonuclease Nicking endonuclease Molecular beacon 

Notes

Acknowledgment

This work was supported by the National Natural Science Foundation of China (Grant 20975113, 31200638, and 21275161).

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Bingjie Zou
    • 1
  • Yinjiao Ma
    • 1
  • Guohua Zhou
    • 1
  1. 1.Department of PharmacologyJinling Hospital, Nanjing University School of MedicineNanjingChina

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