Transcription
Next Generation SequencingGenomics& Medicinehttp://biochem118.stanford.edu/Doug Brutlag, Professor Emeritusof Biochemistry & Medicine (by courtesy)Stanford University School of Medicine Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Illumina Solexa Sequencing Technology Doug Brutlag 2015
Life Sciences 454 Process Overview1) Prepare Adapter Ligated ssDNA Library2) Clonal Amplif icationon 28 µ beads3) Load beads and enzymesin PicoTiter Plate 4) Perform Sequencing by synthesison the 454 Instrument Doug Brutlag 2015
Flowgrams and BaseCallingFlow Order4-merTACG3-mer2-mer1-mer Doug Brutlag 2015
Ion Torrent Sequencinghttp://www.iontorrent.com/ Doug Brutlag 2015
Ion Torrent Sequencinghttp://www.iontorrent.com/ Doug Brutlag 2015
Ion Torrent Sequencinghttp://www.iontorrent.com/ Doug Brutlag 2015
Pacifc Biosciences SMRT Sequencing Doug Brutlag 2015
New PacBio Sequencing Technology t-technology/ Doug Brutlag 2015
New PacBio Sequencing Technology t-technology/ Doug Brutlag 2015
Pacifc Biosciences Sequencing Doug Brutlag 2015
Phospholinked Fluorophores Doug Brutlag 2015
Processive Synthesis Doug Brutlag 2015
Synthesis of Long Duplex DNA Doug Brutlag 2015
Highly Parallel Optics System Doug Brutlag 2015
Circular Templates Gives RedundantSequencing and Accuracy Doug Brutlag 2015
Circular Templates Gives RedundantSequencing and Accuracy Doug Brutlag 2015
The Human GenomeHow fast is the cost going down? 2006: 50 million2008: 500,0002009: 50,0002010: 20,0002011: 5,0002012: 4,0002013: 3,0002014 1,4002015 1,000Thanks to SerafimBrutlagBatzoglou Doug2015
Cost per Raw Megabase ofDNA Sequence Doug Brutlag 2015
Cost per Human Genome Doug Brutlag 2015
Centers for Mendelian Genomicshttp://mendelian.org/ Doug Brutlag 2015
Centers for Mendelian Genomicshttp://mendelian.org/ Doug Brutlag 2015
Genomics Englandhttp://www.genomicsengland.co.uk/ Doug Brutlag 2015
NIH Precision Medicine Initiativehttp://www.nih.gov/precisionmedicine/ Doug Brutlag 2015
Rapid Disease Diagnosis in New BornsScience Translational Medicine 4, 154ra135 ra135.full.html Doug Brutlag 2015
New Gene for Palmoplantar Punktate Keratosishttp://www.nature.com/ng/ Doug Brutlag 2015
Journals PublishingGenomic Medicine Results Nature GeneticsScience Translational MedicineGenome ResearchNew England Journal of MedicineJournal of the American Medical Association Doug Brutlag 2015
Emulsion Based Clonal AmplifcationSingle test tube generation of millions of clonally amplif ied sequencing templatesNo cloning and colony pickingA PCR Reagents Emulsion OilBMicro-reactorsAdapter carryinglibrary DNAAnneal DNA templateto capture beadsBreak micro-reactorsIsolate DNA containingbeads“Water-in-oil”emulsionPerform emulsionPCR Doug Brutlag 2015
Depositing DNA Beads into the PicoTiter PlateLoad Enzyme BeadsLoad beads into PicoTiter PlateCentrifuge Step44 m 70x75mm array of fused optical f ibers with etched wells 1.6 million wells monitored optically through f iber Doug Brutlag 2015
Next Generation Sequencing Doug Brutlag, Professor Emeritus of Biochemistry & Medicine (by courtesy) Stanford University School of Medicine . Single test tube generation of millions of clonally amplif ed sequencing templates No cloning and colony picking "Water-in-oil" emulsion PCR Reagents Emulsion Oil Perform emulsion PCR A B
