Chapter 9Digital Analysis of Genomes9-1
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Sectionsto study9.1 Sequence-specificDNA fragmentation9.2CloningfragmentsofDNA9.3Hybridization9.4The polymerase chain reaction (PCR)9.5DNA sequenceanalysis9-2
9-2 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Sections to study Sections to study 9.1 Sequence 9.1 Sequence-specific DNA fragmentation specific DNA fragmentation 9.2 Cloning fragments of DNA 9.2 Cloning fragments of DNA 9.3 Hybridization 9.3 Hybridization 9.4 The polymerase chain reaction (PCR) 9.4 The polymerase chain reaction (PCR) 9.5 DNA sequence analysis 9.5 DNA sequence analysis 9.6 Bioinformatics: Informati 9.6 Bioinformatics: Information technology and genomes on technology and genomes 9.7 The hemoglobin genes: A comprehensive example 9.7 The hemoglobin genes: A comprehensive example
Tools of modern molecular analysisRecombinant DNA technology:atechnological revolution in themid-1970s.9-3
9-3 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Tools of modern molecular analysis Tools of modern molecular analysis Recombinant DNA technology Recombinant DNA technology: a technological revolution in the technological revolution in the mid-1970s
PlasmiddonorGeneOdonorRecombinantDNAtechnology: alsolationofbacterialplasmidtechnological revolutionin the mid-1970s.PlasmidopenedCIsolatedwithrestrictiongeneMolecularbiologyenzymeBiotechnologyDNAligasebindsendstogetherRecombinantDNAmoleculeTransformationoffreshbacteriumChromasomeODanorgeneTranscriptionTranslationmRNAProteinproduct
9-4 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Recombinant DNA Recombinant DNA technology technology: a technological revolution technological revolution in the mid in the mid-1970s. Molecular biology Molecular biology Biotechnology Biotechnology
Five basic operations in recombinant DNA technologyFragmentlong DNA into smallerfragments and separatethemthroughgelelectrophoresis.2Isolate, amplify, and purify the fragments through molecularcloning.3Use purified DNA fragments as probes toidentify similarsequences in DNA libraries by DNA (or RNA)hybridization.Rapidlyisolate and amplifyknown DNAorRNA sequencesthroughpolymerase chain reaction (PCR).5Determine the nucleotide seguence of DNA through DNAsequencing9-5
9-5 Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display Five basic operations in recombinant DNA technology Five basic operations in recombinant DNA technology 1. Fragment long DNA into smaller fragments and separate Fragment long DNA into smaller fragments and separate them through gel electrophoresis. them through gel electrophoresis. 2. Isolate, amplify, and purify the fragments through molecular Isolate, amplify, and purify the fragments through molecular cloning. cloning. 3. Use purified DNA fragments as probes to identify similar Use purified DNA fragments as probes to identify similar sequences in DNA libraries by sequences in DNA libraries by DNA (or RNA) hybridization. DNA (or RNA) hybridization. 4. Rapidly isolate and amplify known DNA or RNA sequences Rapidly isolate and amplify known DNA or RNA sequences through polymerase chain reaction (PCR). through polymerase chain reaction (PCR). 5. Determine the nucleotide sequence of DNA through DNA Determine the nucleotide sequence of DNA through DNA sequencing. sequencing