1. 04 DNA sequencing

2. The first generation
The Sanger dideoxy method of DNA sequencing

3. The chain termination method3

4. The Sanger dideoxy method of DNA sequencing

6. 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.

7. 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.

8. The second generation: sequencing-by-synthesis
Pyrosequencing
($1.5million, 2008)
Illumina
($48,000 ?, 2010)
SOLiD

9. pyrosequencing 9

10. control Enzyme mix 5ul Substrare mixture 5ul 100pmol primer 1ul
500fmol template 5ul
10xBuffer ul
水 ul
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)

11. Roche/454 FLX Pyrosequencer

15. The third generation: direct measuremeant or synthesis-free
Nanopore

16. Nanopore detector 16

17. 17

18. 18

19. 19