1. DNA sequencing

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

3. The chain termination method

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

10. control Enzyme mix 5ul Substrare mixture 5ul 100pmol primer 1ul 500fmol template 5ul 10xBuffer 10ul 水 74ul GCAGGATG 間隔2min dNTP 0.2x S E A G T C

11. Roche/454 FLX Pyrosequencer

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

16. Nanopore detector