Lecture_11_genome_sequencing

Next gen sequencing plaxorms 454roche gs 20flx 2005

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Unformatted text preview: drai of the human genome was published in Science. 2001 A drai of the human genome was published in Nature. 50,000,000 2002 454 Life Sciences comes out with a pyrosequencing machine. 100,000,000 2008 Next genera=on sequencing machines arrive. Huge 2011 Oxford Nanopore: 600 Million base pairs per hour. Robert Holley and team in 1965 Watson and Crick Messing: World’s most- cited scien=st Francis and Collins: Private Human Genome project. Next- Gen Sequencing PlaXorms 454/Roche GS- 20/FLX (2005) PacBio RS (2009- 2010) 3rd genera=on? Illumina HISeq (2007) 23 Comparison of PlaXorms Technology Reads per run Average Read Length 454 (Roche) 400,000 250- 1000bp SoLID (ABI) 88- 132 Million 35bp bp per run 70 Million Types of errors Subs=tu=on 1 Billion Illumina HISeq 150 Million 100 – 200bp 15 Billion Subs=tu=on with exponen=al increase PacBio 1000- 2000bp 45 Million Inser=ons and dele=ons 45,000 \ Sequencing Methods and Terminology Sanger Sequencing •  The key principle of the Sanger method was the dideoxynucleo=de triphosphates (ddNTPs) as DNA chain terminators. •  These ddNTPs will also be radioac=vely for detec=on in automated sequencing machines. •  Posi=ves: longer reads (600 to 1000 bp). •  Nega=ves: poor coverage (6x), expensive, inaccurate. •  S=ll commonly used for small scale sequencing. Sanger Sequencing Video Sanger Sequencing DNA target sample SHEAR Sanger Sequencing DNA target sample A T C G SHEAR A T C G Close each fragment many times. A...
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