Single-base-resolution and long-coverage sequencing based on single-molecule nanomanipulation

Date

2007

Authors

An, H.
Huang, J.
Lu, M.
Li, X.
Lu, J.
Li, H.
Zhang, Y.
Li, M.
Hu, J.

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Journal article

Citation

Nanotechnology, 2007; 18(22):225101-

Statement of Responsibility

Hongjie An,Jiehuan Huang, Ming Lü, Xueling Li, Junhong Lü, Haikuo Li, Yi Zhang, Minqian Li and Jun Hu

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Abstract

We show new approaches towards a novel single-molecule sequencing strategy which consists of high-resolution positioning isolation of overlapping DNA fragments with atomic force microscopy (AFM), subsequent single-molecule PCR amplification and conventional Sanger sequencing. In this study, a DNA labelling technique was used to guarantee the accuracy in positioning the target DNA. Single-molecule multiplex PCR was carried out to test the contamination. The results showed that the two overlapping DNA fragments isolated by AFM could be successfully sequenced with high quality and perfect contiguity, indicating that single-base resolution and long-coverage sequencing have been achieved simultaneously.

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© 2007 IOP Publishing Ltd

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