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NGS Technologies: Illumina
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Anton Nekrutenko
January 18, 2017
Education
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3k
NGS Technologies: Illumina
Slides describing the principle behind Illumina technology
Anton Nekrutenko
January 18, 2017
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Transcript
Illumina Illumina: Chemistry, Molecular Biology and Data (2018)
Requirements ‣ Reversible termination ‣ Differentiation of nucleotides ‣ Ability
to manipulate on solid support
Preliminary studies
Nucleotides Bentley et al. 2008
Solid support Fedurco et al. 2006
How it works
The overall idea
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Metzker et al. 2010
Extending to paired ends
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Multiplexing
Detection system G,T A,C
How it does not work
Errors
Phasing and Pre-phasing
Illumina chastity filter
Sequence Specific Errors Nakamura et al. 2011
Schematic representation of the (a) inverted repeat and (b) enzyme
preference for the SSE hypothetical mechanistic models. Nakamura K et al. Nucl. Acids Res. 2011;39:e90-e90 © The Author(s) 2011. Published by Oxford University Press.
A typical run Minoche et al. 2011
Coverage as function of GC% Minoche et al. 2011
Low quality bases and alignment Minoche et al. 2011
Context dependency Minoche et al. 2011
None
True seq PCR free