04 DNA sequencing

The first generation The Sanger dideoxy method of DNA sequencing

The chain termination method 3

The Sanger dideoxy method of DNA sequencing

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 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 ($1.5million, 2008) Illumina ($48,000 ?, 2010) SOLiD

pyrosequencing 9

control Enzyme mix 5ul Substrare mixture 5ul 100pmol primer 1ul 500fmol template 5ul 10xBuffer 10ul 水 74ul GCAGGATG 間隔2min dNTP 0.2x (1,133,379) (904,103) 969,984 667,372 54,249 (253,798) (244,154) (253,619) (199,549) (236,731) 196,470 (163,395) (58,632) (47,684) S E A G T C (108)

Roche/454 FLX Pyrosequencer

The third generation: direct measuremeant or synthesis-free Nanopore

Nanopore detector 16

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