DNA sequencing The first generation The Sanger dideoxy

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DNA sequencing

DNA sequencing

The first generation The Sanger dideoxy method of DNA sequencing

The first generation The Sanger dideoxy method of DNA sequencing

The chain termination method

The chain termination method

The Sanger dideoxy method of DNA sequencing

The Sanger dideoxy method of DNA sequencing

l l This method was costly, at around $10 per base pair in 1985,

l l This method was costly, at around $10 per base pair in 1985, but the development of automated sequencing systems and advancements in technology reduced the price to $1 per base by 1995, and allowed sequencing of up to 100, 000 bases per day. The cost dropped further to $0. 10 per base in 1998 with the development of the ABI Prism sequencer, which made it possible toundertake larger-scale sequencing projects. • In the context of highthroughput shotgun genomic sequencing, Sanger sequencing costs on the order of $0. 50 per kilobase.

In search of the $1000 genome • National Institutes of Health (NIH), specifically the

In search of the $1000 genome • National Institutes of Health (NIH), specifically the NIH’s National Human Genome Research Institute (NHGRI), has actively sponsored researchers with $99. 5 million of external funding to develop sequencing technologies since the completion of the Human Genome Project. • $100, 000 initially and then as little as $1000.

The second generation: sequencing-by-synthesis • Pyrosequencing ($1. 5 million, 2008) • Illumina ($48, 000

The second generation: sequencing-by-synthesis • Pyrosequencing ($1. 5 million, 2008) • Illumina ($48, 000 ? , 2010) • SOLi. D

pyrosequencing

pyrosequencing

control Enzyme mix 5 ul Substrare mixture 5 ul 100 pmol primer 1 ul

control Enzyme mix 5 ul Substrare mixture 5 ul 100 pmol primer 1 ul 500 fmol template 5 ul 10 x. Buffer 10 ul 水 74 ul GCAGGATG 間隔 2 min d. NTP 0. 2 x (1, 133, 379) (904, 103 ) 969, 984 667, 372 54, 249 (244, 154) (58, 632) (108) S (47, 684) E 196, 470 A (163, 395) G (253, 798) (199, 549) T (253, 619) (236, 731 ) C

Roche/454 FLX Pyrosequencer (high-throughput sequencing)

Roche/454 FLX Pyrosequencer (high-throughput sequencing)

The third generation: direct measuremeant or synthesis-free • Nanopore

The third generation: direct measuremeant or synthesis-free • Nanopore

Nanopore detector

Nanopore detector

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