Duplex Sequencing
Duplex Sequencing
Duplex sequencing is a tag-based error correction method to improve sequencing accuracy . In this method, adapters (with primer sequences and random 12 bp indices) are ligated onto the template and amplified using PCR. Deep sequencing provides consensus sequence information from every unique molecular tag. Based on molecular tags and sequencing primers, duplex sequences are aligned, determining the true sequence on each DNA strand.
Pros:
- Very low error rate due to duplex tagging system
- PCR amplification errors can be detected and removed from analysis
- No additional library preparation steps after addition of adapters
Cons:
- PCR amplification errors
- Non-linear PCR amplification can lead to biases affecting reproducibility
- PCR biases can underrepresent GC-rich templates
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- Wang Y., Waters J., Leung M. L., Unruh A., Roh W., et al. (2014) Clonal evolution in breast cancer revealed by single nucleus genome sequencing. Nature 512: 155-160
- Kennedy S. R., Salk J. J., Schmitt M. W. and Loeb L. A. (2013) Ultra-sensitive sequencing reveals an age-related increase in somatic mitochondrial mutations that are inconsistent with oxidative damage. PLoS Genet 9: e1003794