Book contents
- Frontmatter
- Contents
- Preface
- Acknowledgments
- Chapter 1 Life Forms and Their Origins
- Chapter 2 Nucleic Acids
- Chapter 3 Proteins
- Chapter 4 Simple Chromosomes
- Chapter 5 Chromosomes of Eukarya
- Chapter 6 Genome Content
- Chapter 7 RNA Synthesis 1: Transcription
- Chapter 8 RNA Synthesis 2: Processing
- Chapter 9 Abundance of RNAs in Bacteria
- Chapter 10 Abundance of RNAs in Eukarya
- Chapter 11 Protein Synthesis
- Chapter 12 DNA Replication
- Chapter 13 Chromosome Replication
- Chapter 14 Molecular Events of Recombination
- Chapter 15 Micromutations
- Chapter 16 Repair of Altered DNA
- Chapter 17 Reproduction of Bacteria
- Chapter 18 Horizontal Gene Transfer in Bacteria
- Chapter 19 Cell Cycles of Eukarya
- Chapter 20 Meiosis
- Chapter 21 Chromosomal Abnormalities
- Chapter 22 Life Cycles of Eukarya
- Chapter 23 Reproduction of Viruses
- Chapter 24 Genetic Processes in Development
- Chapter 25 Sex Determination and Dosage Compensation
- Chapter 26 Cancer
- Chapter 27 Cutting, Sorting, and Copying DNA
- Chapter 28 Genotyping by DNA Analysis
- Chapter 29 Genetically Engineered Organisms
- Chapter 30 Genomics
- Chapter 31 Behavior of Genes and Alleles
- Chapter 32 Probability and Statistics Toolkit
- Chapter 33 Genes, Environment, and Interactions
- Chapter 34 Locating Genes
- Chapter 35 Finding and Detecting Mutations
- Chapter 36 Cytoplasmic Inheritance
- Chapter 37 Genetic Variation in Populations
- Chapter 38 Mutation, Migration, and Genetic Drift
- Chapter 39 Natural Selection
- Chapter 40 Quantitative Genetics
- Chapter 41 Speciation
- Chapter 42 Molecular Evolution and Phylogeny
- Glossary
- Index
Chapter 27 - Cutting, Sorting, and Copying DNA
Published online by Cambridge University Press: 05 June 2012
- Frontmatter
- Contents
- Preface
- Acknowledgments
- Chapter 1 Life Forms and Their Origins
- Chapter 2 Nucleic Acids
- Chapter 3 Proteins
- Chapter 4 Simple Chromosomes
- Chapter 5 Chromosomes of Eukarya
- Chapter 6 Genome Content
- Chapter 7 RNA Synthesis 1: Transcription
- Chapter 8 RNA Synthesis 2: Processing
- Chapter 9 Abundance of RNAs in Bacteria
- Chapter 10 Abundance of RNAs in Eukarya
- Chapter 11 Protein Synthesis
- Chapter 12 DNA Replication
- Chapter 13 Chromosome Replication
- Chapter 14 Molecular Events of Recombination
- Chapter 15 Micromutations
- Chapter 16 Repair of Altered DNA
- Chapter 17 Reproduction of Bacteria
- Chapter 18 Horizontal Gene Transfer in Bacteria
- Chapter 19 Cell Cycles of Eukarya
- Chapter 20 Meiosis
- Chapter 21 Chromosomal Abnormalities
- Chapter 22 Life Cycles of Eukarya
- Chapter 23 Reproduction of Viruses
- Chapter 24 Genetic Processes in Development
- Chapter 25 Sex Determination and Dosage Compensation
- Chapter 26 Cancer
- Chapter 27 Cutting, Sorting, and Copying DNA
- Chapter 28 Genotyping by DNA Analysis
- Chapter 29 Genetically Engineered Organisms
- Chapter 30 Genomics
- Chapter 31 Behavior of Genes and Alleles
- Chapter 32 Probability and Statistics Toolkit
- Chapter 33 Genes, Environment, and Interactions
- Chapter 34 Locating Genes
- Chapter 35 Finding and Detecting Mutations
- Chapter 36 Cytoplasmic Inheritance
- Chapter 37 Genetic Variation in Populations
- Chapter 38 Mutation, Migration, and Genetic Drift
- Chapter 39 Natural Selection
- Chapter 40 Quantitative Genetics
- Chapter 41 Speciation
- Chapter 42 Molecular Evolution and Phylogeny
- Glossary
- Index
Summary
Overview
Because the genes of all organisms consist of DNA, and because a gene's structure largely determines its functional properties, analysis of any particular gene demands the isolation and structural analysis of DNA. To manipulate DNA experimentally, or even to determine its base sequence, chromosomal DNA must be reduced to small fragments, which are then sorted, identified, copied, and stored for future use.
This chapter explains some basic principles of cutting DNA enzymatically into fragments, separating pieces of DNA or RNA by size, making DNA copies of RNA, identifying one particular piece of DNA or RNA, copying DNA, and establishing large collections of DNA fragments for storage and retrieval.
Cutting and Fractionating DNA
Restriction Enzymes
It is easy to isolate chromosomal DNA. However, chromosomes tend to be long, and, even after random breakage, the DNA isolated from chromosomes exists as intractably long pieces. It is convenient to cut DNA into appropriate-sized fragments with bacterial restriction endonucleases (often simply called restriction enzymes), which cut only at particular sites.
Bacteria protect themselves against viral infection by cutting foreign DNA with restriction enzymes, so called because they restrict foreign DNA's ability to invade the cell. The cell distinguishes its own DNA from foreign DNA by methylating certain bases shortly after replication, using modification enzymes. At least one strand of the bacterium's own DNA is modified and therefore resistant to restriction, whereas foreign DNA is not modified and therefore susceptible to restriction.
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- Information
- Fundamental Genetics , pp. 254 - 264Publisher: Cambridge University PressPrint publication year: 2004