Marzillier101207

Marzillier101207 - Genome Sequencing Technologies Jutta...

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Jutta Marzillier, 2007 Genome Sequencing Technologies Jutta Marzillier, Ph.D. Lehigh University Department of Biological Sciences Iacocca Hall
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Jutta Marzillier, 2007 Sciences start with Observation
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Jutta Marzillier, 2007 Sciences start with Observation and flourish with Introduction of Technology
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Jutta Marzillier, 2007 Life Sciences Thrive Trough the Co-laboration between Biologists and Engineers Biologist BioMedical/ Chemical/ Mechanical Engineer Computer Engineer/ Bioinformatics
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Jutta Marzillier, 2007 Components that lead to completion of the Human Genome Project Life Sciences : Discovery of DNA (1953) Invention of DNA Sequencing (1975) Polymerase Chain Reaction (PCR, 1985) Human Genome published (2003) Engineering : Dye coupling to DNA (1986) Capillary electrophoresis (1995) Robots Automated Sequencer (1986) Microfluidics Nanotechnology Computer Sciences/ Bioinformatics : Computer languages (e.g. FORTRAN 1956) Floppy disc (1970), hard drives 1 st sequence database (EMBL) opens (1980) Genbank (1982) BLAST (Basic Local Alignment Search Tool) (1990) World Wide Web (1989) ‘PC’ revolution (1 st PC by IBM in 1981)
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Jutta Marzillier, 2007 What is the Genome ? The human body has about 100 trillion cells Each cell carries the same genetic information ( genome ) in its nucleus Depending on the cellular context only a subset of genes is expressed ( transcriptome ) © 2002 The Center for the Advancement of Genomics (TCAG). Why study genes? The study of genes is crucial to our understanding of the cell, development and disease.
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Jutta Marzillier, 2007 Main Biological Dogma protein Expressed proteins in particular cellular context Translation and post- translational modifications Proteomics DNA mRNA ‘working library’: coding, non-coding, regulatory elements Gene expression for particular cellular context Transcription Genomics Transcriptomics RNA
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Jutta Marzillier, 2007 DNA structure and synthesis DNA: chemically linked chain of nucleotides which each consists of a phosphate, a sugar (ribose), and a nucleobase ( A denine, G uanine, C ytosine, T hymine)
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Jutta Marzillier, 2007 DNA sequencing – Principle of DNA synthesis ± Amplification of targeted DNA strand in presence of - primer that only hybridizes to one complementary strand- -(DNA) polymerase , (or Taq) - deoxynucleotides (dATP,dTTP,dCTP,dGTP).
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Jutta Marzillier, 2007 Frederick Sanger * 13 August 1918 Two time Nobel laureate in chemistry 1958: identification of the amino acid sequence of insulin 1980: developed the chain termination method for DNA sequencing
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Jutta Marzillier, 2007 Dideoxy Method of Sequencing (Sanger, 1975) ± DNA synthesis is carried out in the presence of limiting amounts of dideoxyribonucleoside triphosphates that results in chain termination ± Original method used radio-labeled primers or dideoxynucleotides
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Jutta Marzillier, 2007 Sanger DNA sequencing using radiolabel
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Jutta Marzillier, 2007 Fluorescently labeled ddNTPs used for
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Marzillier101207 - Genome Sequencing Technologies Jutta...

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