Talk outline:
•Structure and chemistry of DNA
•Development of classical sequencing technologies and the human genome project
•Next generation sequencing platform and technology
If time, some applications of NGS
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DNA first identified (nuclein) by Friedrich Miescher in 1869
Chemical makeup by Phoebus Levene 1909-1919, and suggested sugar-phosphate
backbone structure
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DNA first identified (nuclein) by Friedrich Miescher in 1869
Chemical makeup by Phoebus Levene 1909-1919, and suggested sugar-phosphate
backbone structure
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Explain complementary strands, directionality, and importance of direction in
interpretation
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Encodes most information about an organism – protein coding regions, regulatory
sequences (but mentioned epigenetics etc).
Knowing the sequence helps understand how biological processes work, and being
able to find changes can help to explain genetic disorders.
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Leroy Hood lab at CalTech
Applied Biosystem 370 sequencer
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2000 first draft genome was announced
2003 complete genome announced
2006 final chromosome published in Nature
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Not only massively parallel, also new sample preparation so doesn’t need specific
primer sites
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Started off as 454, launched in 2005 as GS20
Talk about iterative models later on
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Don’t forget to talk about runs of the same base
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Expected further upgrade to 700bp reads
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Started off as Solexa, launched in 2006
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Not strictly true now – use Y-shaped adapters
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Immobilisation – single cycle
Isothermal bridge amplification
~2000 copies per cluster
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Explain unpredictability of developments
Now v1.7 software. 3-fold increase in data yield just by software improvements for
base calling
New hardware – just increase area – 100Gbp/run per flowcell
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This time last year, maximum yield/day was 2Gbp. Now 25Gbp
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Started off as Agencourt, bought by AB in 2006, and launched in 2007
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