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NGS Technologies: Illumina
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Anton Nekrutenko
January 18, 2017
Education
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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
None
None
None
None
None
None
None
None
None
None
None
Metzker et al. 2010
Extending to paired ends
None
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