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Gold nanoparticle-assisted primer walking for closing the human chromosomal gap

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  • H Li, Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Science, Kina
  • B Shi, Institute of Biophysics, Chinese Academy of Sciences, Beijing, Kina
  • X Li, Shanghai Center for Bioinformation Technology, Danmark
  • Steffen Nyegaard, Danmark
  • M Lü, Shanghai JiaoTong University, Kina
  • J Lü, Chinese Academy of Sciences, Kina
  • X Cao, College of Life Science and Biotechnology, Shanghai JiaoTong University, Shanghai, Kina
  • Q Huang, Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Kina
  • C Fan, Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Kina
  • R. Chen, Institute of Biophysics, Chinese Academy of Sciences, Kina
  • J Hu, Laboratory of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Kina
The finished sequence of the human genome still contains 260 euchromatic gaps. All the PCR-based genome walking techniques used to close gaps have common limitations, such as low efficiency and low specificity. We herein describe a strategy to solve this problem by employing gold nanoparticles (AuNPs) to improve the efficiency in primer walking amplification. We used this strategy to close a gap in human chromosome 5 containing a DNA stretch composed of the 12SAT repeat. The obtained gap sequence is highly conserved among several mammalian genomes. The demonstrated AuNP-assisted primer walking strategy is capable of effectively improving the specificity of PCR amplification and enriching the yield of target DNA fragments; it offers a new avenue for closing gaps left by current sequencing methods
OriginalsprogEngelsk
TidsskriftAnalytical Methods
Vol/bind5
Nummer18
Sider (fra-til)4746-4749
Antal sider4
ISSN1759-9660
DOI
StatusUdgivet - 21 sep. 2013

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